903 results on '"AMP Deaminase"'
Search Results
2. Adenosine monophosphate deaminase 3 null mutation causes reduction of naive T cells in mouse peripheral blood
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Jin Huk Choi, Sara Ludwig, Bruce Beutler, Xue Zhong, Miao Tang, Priscilla Anderton, Lijing Su, Evan Nair-Gill, Xiaoming Zhan, Jamie Russell, Xiaohong Li, Thomas Gallagher, and Jianhui Wang
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Mice, Knockout ,Adenosine monophosphate ,Mutation ,Immunobiology and Immunotherapy ,T-Lymphocytes ,Lymphocyte ,Spleen ,AMP deaminase ,Hematology ,Biology ,medicine.disease_cause ,Molecular biology ,Adenosine Monophosphate ,AMP Deaminase ,Mice ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Loss of Function Mutation ,Knockout mouse ,medicine ,Animals ,Adenosine triphosphate ,CD8 - Abstract
Adenosine monophosphate deaminase 3 (Ampd3) encodes the erythrocyte isoform of the adenosine monophosphate (AMP) deaminase gene family. Mutations in this gene have been reported in humans, leading to autosomal-recessive erythrocyte AMP deaminase deficiency. However, the mutation is considered clinically asymptomatic. Using N-ethyl-N-nitrosourea mutagenesis to find mutations that affect peripheral lymphocyte populations, we identified 5 Ampd3 mutations (Ampd3guangdong, Ampd3carson, Ampd3penasco, Ampd3taos, and Ampd3commanche) that strongly correlated with a reduction in naive CD4+ T and naive CD8+ T-cell populations. Causation was confirmed by targeted ablation of Ampd3. Knockout mice had reduced frequencies of CD62LhiCD44lo CD4+ naive and CD8+ naive T cells. Interestingly, these phenotypes were restricted to T cells circulating in peripheral blood and were not seen in T cells from secondary lymphoid organs (lymph nodes and spleen). We found that reduction of naive T cells in the peripheral blood of Ampd3−/− mice was caused by T-cell–extrinsic factor(s), which we hypothesize to be elevated levels of adenosine triphosphate released by Ampd3-deficient erythrocytes. These findings provide an example in which disruption of an erythrocyte-specific protein can affect the physiological status of lymphocytes in peripheral blood.
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- 2020
3. Valine acts as a nutritional signal in brain to activate TORC1 and attenuate postprandial ammonia-N excretion in Chinese perch (Siniperca chuatsi)
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Xu-Fang Liang, Jie Wang, Ping Ren, Jiao Li, Yanpeng Zhang, Kang Huang, Shan He, and Lin-Jie Shi
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Fish Proteins ,medicine.medical_specialty ,Physiology ,Protein metabolism ,Mechanistic Target of Rapamycin Complex 1 ,Aquatic Science ,Protein degradation ,Biology ,Biochemistry ,AMP Deaminase ,Excretion ,03 medical and health sciences ,chemistry.chemical_compound ,Glutamate Dehydrogenase ,Ammonia ,Valine ,Internal medicine ,medicine ,Animals ,Injections, Intraventricular ,030304 developmental biology ,chemistry.chemical_classification ,Ribosomal Protein S6 ,0303 health sciences ,Muscles ,Glutamate dehydrogenase ,Brain ,04 agricultural and veterinary sciences ,General Medicine ,Postprandial Period ,Amino acid ,Endocrinology ,Postprandial ,Liver ,chemistry ,Perches ,Ribosomal protein s6 ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,Signal Transduction - Abstract
An emerging concept is that the hypothalamic nutrient sensor can regulate peripheral energy metabolism via a brain-liver circuit. Valine is an essential branched-chain amino acid (BCAA) that drives intracellular signaling cascades by the activation of target of rapamycin complex 1 (TORC1) which is critical to protein metabolism in mammals. However, in teleost fish, it remains scarce in this area especially about how the intraventricular (ICV) injection of valine can mediate the protein metabolism in peripheral organs. This study would tentatively explore the effects of ICV injection of valine on protein metabolism in peripheral organs through evaluating the postprandial ammonia-N excretion rate in Chinese perch. The control group was injected with 5-μL PBS, and the Val group was injected with 20-μg L valine dissolved into 5-μL PBS. The ammonia-N excretion rate of Val group was lower than control group at 4-, 12-, and 24-h postinjection, while the concertation of plasma glucose was increased sharply at 0.5-, 4-, 12-, and 24-h postinjection. We further checked both mRNA level and the enzyme activity of glutamate dehydrogenase (GDH) in the liver and adenosine monophosphate deaminase (AMPD) in muscle, and we found that they were obviously decreased in Val group at 4-, 12-, and 24-h postinjection. The phosphorylation level of ribosomal protein S6, a downstream target protein of TORC1, was markedly enhanced in the liver of Val group at 4-, 12-, and 24-h postinjection. Collectively, these results illustrated that ICV injection of valine can attenuate protein degradation in peripheral organs by depressing the GDH and AMPD enzyme activity; on the other hand, the injected valine can trigger the activation of TORC1 in the liver via a brain-liver circuit and then interdict proteolysis.
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- 2020
4. Modelling of energy metabolism and analysis of pH variations in postmortem muscle
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Chengcheng Wang, Jinglu Tan, Sulaiman K. Matarneh, and David E. Gerrard
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Male ,Models, Statistical ,biology ,Chemistry ,Kinetics ,Sus scrofa ,Adenylate kinase ,AMP deaminase ,Mitochondrion ,Hydrogen-Ion Concentration ,Phosphofructokinase activity ,Mitochondria ,ATP hydrolysis ,Biophysics ,biology.protein ,Pork Meat ,Animals ,Glycolysis ,Creatine kinase ,Female ,Energy Metabolism ,Muscle, Skeletal ,Food Science - Abstract
A kinetic model structure was developed to describe the major variations in energy metabolism and to gain further understanding of pH changes in postmortem muscle experimentally observed with an in vitro glycolytic system. Comparison with experiments showed that the model could describe the kinetics of major metabolites and pH under varied conditions. Optimized model parameters definitively and consistently showed the observed effects of mitochondria, indicating a desirable level of model complexity. Simulation and analysis of pH variations based on the model suggested that phosphofructokinase activity has the strongest impact on the rate and extent of postmortem pH decline. Postmortem pH is also influenced by rates of ATP hydrolysis and glycolysis, and to a much lesser extent, pH buffering capacity. Other reactions, including those mediated by creatine kinase, adenylate kinase, and AMP deaminase, have minimal effects on postmortem pH decline.
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- 2021
5. Biochemical and physiological investigations on adenosine 5ʹ monophosphate deaminase fromPlasmodium spp
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Dipti Singh, Kota Arun Kumar, Hemalatha Balaram, Sandeep Dey, and Lakshmeesha Kempaiah Nagappa
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Adenosine monophosphate ,Plasmodium falciparum ,Allosteric regulation ,Protozoan Proteins ,Adenylate kinase ,Biology ,Microbiology ,Catalysis ,AMP Deaminase ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Adenosine Triphosphate ,Allosteric Regulation ,Protein-fragment complementation assay ,Animals ,Humans ,Malaria, Falciparum ,Energy charge ,Molecular Biology ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,030306 microbiology ,Kinase ,AMP deaminase ,Adenosine Monophosphate ,Mice, Inbred C57BL ,Enzyme ,chemistry ,Biochemistry - Abstract
The interplay between ATP generating and utilizing pathways in a cell is responsible for maintaining cellular ATP/energy homeostasis that is reflected by Adenylate Energy Charge (AEC) ratio. Adenylate kinase (AK), that catalyzes inter-conversion of ADP, ATP and AMP, plays a major role in maintaining AEC and is regulated by cellular AMP levels. Hence, the enzymes AMP deaminase (AMPD) and nucleotidases, which catabolize AMP, indirectly regulate AK activity and in-turn affect AEC. Here, we present the first report on AMPD from Plasmodium, the causative agent of malaria. The recombinant enzyme expressed in Saccharomyces cerevisiae was studied using functional complementation assay and residues vital for enzyme activity have been identified. Similarities and differences between Plasmodium falciparum AMPD (PfAMPD) and its homologs from yeast, Arabidopsis and humans are also discussed. The AMPD gene was deleted in the murine malaria parasite P. berghei and was found to be dispensable during all stages of the parasite life cycle. However, when episomal expression was attempted, viable parasites were not obtained, suggesting that perturbing AMP homeostasis by over-expressing AMPD might be lethal. As AMPD is known to be allosterically modulated by ATP, GTP and phosphate, allosteric activators of PfAMPD could be developed as anti-parasitic agents.
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- 2019
6. AMP deamination is sufficient to replicate an atrophy-like metabolic phenotype in skeletal muscle
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Spencer G. Miller, Jeffrey J. Brault, Andrew S. Law, Thomas M. O’Connell, Paul S. Hafen, David L. Logsdon, Catherine B. Springer, and Carol A. Witczak
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medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Article ,AMP Deaminase ,Mice ,Endocrinology ,Adenosine Triphosphate ,AMP-activated protein kinase ,Internal medicine ,medicine ,Myocyte ,Animals ,Glycolysis ,Muscle, Skeletal ,biology ,Chemistry ,Skeletal muscle ,AMPK ,AMP deaminase ,Muscle atrophy ,Adenosine Monophosphate ,Adenosine Diphosphate ,Muscular Atrophy ,medicine.anatomical_structure ,Phenotype ,Mitochondrial biogenesis ,Deamination ,biology.protein ,medicine.symptom - Abstract
Background Skeletal muscle atrophy, whether caused by chronic disease, acute critical illness, disuse or aging, is characterized by tissue-specific decrease in oxidative capacity and broad alterations in metabolism that contribute to functional decline. However, the underlying mechanisms responsible for these metabolic changes are largely unknown. One of the most highly upregulated genes in atrophic muscle is AMP deaminase 3 (AMPD3: AMP → IMP + NH3), which controls the content of intracellular adenine nucleotides (AdN; ATP + ADP + AMP). Given the central role of AdN in signaling mitochondrial gene expression and directly regulating metabolism, we hypothesized that overexpressing AMPD3 in muscle cells would be sufficient to alter their metabolic phenotype similar to that of atrophic muscle. Methods AMPD3 and GFP (control) were overexpressed in mouse tibialis anterior (TA) muscles via plasmid electroporation and in C2C12 myotubes using adenovirus vectors. TA muscles were excised one week later, and AdN were quantified by UPLC. In myotubes, targeted measures of AdN, AMPK/PGC-1α/mitochondrial protein synthesis rates, unbiased metabolomics, and transcriptomics by RNA sequencing were measured after 24 h of AMPD3 overexpression. Media metabolites were measured as an indicator of net metabolic flux. At 48 h, the AMPK/PGC-1α/mitochondrial protein synthesis rates, and myotube respiratory function/capacity were measured. Results TA muscles overexpressing AMPD3 had significantly less ATP than contralateral controls (−25%). In myotubes, increasing AMPD3 expression for 24 h was sufficient to significantly decrease ATP concentrations (−16%), increase IMP, and increase efflux of IMP catabolites into the culture media, without decreasing the ATP/ADP or ATP/AMP ratios. When myotubes were treated with dinitrophenol (mitochondrial uncoupler), AMPD3 overexpression blunted decreases in ATP/ADP and ATP/AMP ratios but exacerbated AdN degradation. As such, pAMPK/AMPK, pACC/ACC, and phosphorylation of AMPK substrates, were unchanged by AMPD3 at this timepoint. AMPD3 significantly altered 191 out of 639 detected intracellular metabolites, but only 30 transcripts, none of which encoded metabolic enzymes. The most altered metabolites were those within purine nucleotide, BCAA, glycolysis, and ceramide metabolic pathways. After 48 h, AMPD3 overexpression significantly reduced pAMPK/AMPK (−24%), phosphorylation of AMPK substrates (−14%), and PGC-1α protein (−22%). Moreover, AMPD3 significantly reduced myotube mitochondrial protein synthesis rates (−55%), basal ATP synthase-dependent (−13%), and maximal uncoupled oxygen consumption (−15%). Conclusions Increased expression of AMPD3 significantly decreased mitochondrial protein synthesis rates and broadly altered cellular metabolites in a manner similar to that of atrophic muscle. Importantly, the changes in metabolites occurred prior to reductions in AMPK signaling, gene expression, and mitochondrial protein synthesis, suggesting metabolism is not dependent on reductions in oxidative capacity, but may be consequence of increased AMP deamination. Therefore, AMP deamination in skeletal muscle may be a mechanism that alters the metabolic phenotype of skeletal muscle during atrophy and could be a target to improve muscle function during muscle wasting.
- Published
- 2021
7. A functional mutation in the AMPD1 promoter region affects promoter activity and breast meat freshness in chicken
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Liao Juan, Wang Gang, X Chen, and Yu Shigang
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0301 basic medicine ,Adenosine monophosphate ,Inosine monophosphate ,animal structures ,Genotype ,Biology ,Protein degradation ,medicine.disease_cause ,Poultry ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,Genetics ,medicine ,Food Quality ,Animals ,Food science ,Promoter Regions, Genetic ,Gene ,Transcription factor ,Alleles ,Mutation ,0402 animal and dairy science ,Purine nucleotide cycle ,food and beverages ,Promoter ,04 agricultural and veterinary sciences ,General Medicine ,040201 dairy & animal science ,030104 developmental biology ,chemistry ,Animal Science and Zoology ,Female ,Chickens - Abstract
Freshness is an important index to determine the quality deterioration (protein degradation and changes in appearance) of chilled chicken meat and is a primary consideration of consumers. Adenosine monophosphate deaminase 1 (AMPD1) catalyzes the deamination of adenosine monophosphate to inosine monophosphate in skeletal muscle and is the rate-limiting step in the purine nucleotide cycle. Inosine monophosphate is regarded as an important indicator of meat freshness in chicken. This study investigated the association of polymorphisms in the chicken AMPD1 promoter region with meat freshness during freezing storage. An SNP (c. -905GA) was found to be associated with the freshness (K-value) of chicken breast meat. Chickens with the AA genotype had significantly lower K-values than those with GG and AG genotypes (P 0.01). Individuals with the AA genotype also had higher breast meat AMPD1 mRNA levels than did those with the GG and AG genotypes (P 0.01, P 0.05). A luciferase assay revealed that genotype AA had greater transcriptional activity than genotype GG. Transcription factor binding site analysis identified distinct putative transcription factor binding sites in the two alleles of mutation site c. -905. In summary, we identified an SNP (c. -905GA) in the promoter region of the AMPD1 gene that may modulate the binding affinity of different transcription factors to control AMPD1 expression and affect the freshness K-value of chicken meat.
- Published
- 2020
8. Metabolic Buffer Analysis Reveals the Simultaneous, Independent Control of ATP and Adenylate Energy Ratios
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James R. Krycer, Jordan Ang, and Edward J. Hancock
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Bioenergetics ,Biomedical Engineering ,Biophysics ,Adenylate kinase ,Bioengineering ,Biochemistry ,Biomaterials ,Synthetic biology ,Mice ,Adenosine Triphosphate ,Control theory ,Animals ,Control (linguistics) ,Creatine Kinase ,Mechanism (biology) ,Chemistry ,Muscles ,AMP deaminase ,Metabolism ,Cell biology ,Metabolic pathway ,Phosphagen ,Life Sciences–Mathematics interface ,Biological regulation ,Energy Metabolism ,Energy (signal processing) ,Biotechnology - Abstract
Determining the underlying principles behind biological regulation is important for understanding the principles of life, treating complex diseases, and creatingde novosynthetic biology. Buffering - the use of reservoirs of molecules to maintain molecular concentrations - is a widespread and important mechanism for biological regulation. However, a lack of theory has limited our understanding of its roles and quantified effects. Here, we study buffering in energy metabolism using control theory and novel buffer analysis. We find that buffering can enable the simultaneous, independent control of multiple coupled outputs. In metabolism, adenylate kinase and AMP deaminase enable simultaneous control of ATP and adenylate energy ratios, while feedback on metabolic pathways is fundamentally limited to controlling one of these outputs. We also quantify the regulatory effects of the phosphagen system - the above buffers and creatine kinase - revealing which mechanisms regulate which outputs. The results are supported by human muscle and mouse adipocyte data. Together, these results illustrate the synergy of feedback and buffering in molecular biology to simultaneously control multiple outputs.
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- 2020
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9. Increased AMP deaminase activity decreases ATP content and slows protein degradation in cultured skeletal muscle
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Joshua S. Morgan, Jeffrey J. Brault, Carol A. Witczak, Patrick R. Davis, Nicolas A. Verhoeven, David A. Tulis, and Spencer G. Miller
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0301 basic medicine ,Adenosine monophosphate ,medicine.medical_specialty ,Proteasome Endopeptidase Complex ,AMP deaminase activity ,Endocrinology, Diabetes and Metabolism ,Muscle Fibers, Skeletal ,030209 endocrinology & metabolism ,Protein degradation ,AMP-Activated Protein Kinases ,Article ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Endocrinology ,Adenosine Triphosphate ,Adenine nucleotide ,Internal medicine ,medicine ,Animals ,Phosphorylation ,Protein kinase A ,Muscle, Skeletal ,Cells, Cultured ,Ubiquitin ,AMP deaminase ,Adenosine Monophosphate ,Adenosine Diphosphate ,Adenosine diphosphate ,Protein Subunits ,030104 developmental biology ,chemistry ,Biochemistry ,Proteolysis ,Adenosine triphosphate - Abstract
Background Protein degradation is an energy-dependent process, requiring ATP at multiple steps. However, reports conflict as to the relationship between intracellular energetics and the rate of proteasome-mediated protein degradation. Methods To determine whether the concentration of the adenine nucleotide pool (ATP + ADP + AMP) affects protein degradation in muscle cells, we overexpressed an AMP degrading enzyme, AMP deaminase 3 (AMPD3), via adenovirus in C2C12 myotubes. Results Overexpression of AMPD3 resulted in a dose- and time-dependent reduction of total adenine nucleotides (ATP, ADP and AMP) without increasing the ADP/ATP or AMP/ATP ratios. In agreement, the reduction of total adenine nucleotide concentration did not result in increased Thr172 phosphorylation of AMP-activated protein kinase (AMPK), a common indicator of intracellular energetic state. Furthermore, LC3 protein accumulation and ULK1 (Ser 555) phosphorylation were not induced. However, overall protein degradation and ubiquitin-dependent proteolysis were slowed by overexpression of AMPD3, despite unchanged content of several proteasome subunit proteins and proteasome activity in vitro under standard conditions. Conclusions Altogether, these findings indicate that a physiologically relevant decrease in ATP content, without a concomitant increase in ADP or AMP, is sufficient to decrease the rate of protein degradation and activity of the ubiquitin-proteasome system in muscle cells. This suggests that adenine nucleotide degrading enzymes, such as AMPD3, may be a viable target to control muscle protein degradation and perhaps muscle mass.
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- 2020
10. Ultrastructural Localization of Histidine-rich Glycoprotein in Skeletal Muscle Fibers: Colocalization With AMP Deaminase
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Alessandra Falleni, Letizia Mattii, Sabina Frascarelli, Francesco Bianchi, Grazia Chiellini, Matilde Masini, Greta Alì, and Antonietta Raffaella Maria Sabbatini
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Adult ,Male ,Histology ,Histidine-rich glycoprotein ,Muscle Fibers, Skeletal ,Intracellular Space ,Sarcomere ,AMP Deaminase ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,030304 developmental biology ,Aged ,chemistry.chemical_classification ,0303 health sciences ,muscle biopsies ,nucleus ,Colocalization ,Skeletal muscle ,Proteins ,AMP deaminase ,immunohistochemistry ,metallochaperone ,sarcomere ,Articles ,Cell biology ,Rats ,Protein Transport ,medicine.anatomical_structure ,chemistry ,030220 oncology & carcinogenesis ,Ultrastructure ,Immunohistochemistry ,Female ,Anatomy ,Glycoprotein - Abstract
Histidine-rich glycoprotein (HRG) is a plasma protein synthesized by the liver. We have given the first evidence of a tissue localization of HRG demonstrating its presence in skeletal muscle, associated with the zinc enzyme AMP deaminase (AMPD1). Moreover, we have shown that muscle cells do not synthesize HRG, but they can internalize it from plasma. We have recently demonstrated by confocal laser scanning microscopy that in human skeletal muscle, HRG is mainly localized in the myofibrils, preferentially at the I-band of the sarcomere, in the sarcoplasm, and in the nuclei. Using transmission electron microscopy and immunogold analysis, we carried out this study on human and rat normal skeletal muscles with the purpose to deepen the ultrastructural localization of HRG in skeletal muscle fibers. The immunogold analysis evidenced the presence of HRG in the sarcomeres, mainly in the I-band and to a less extent in the A-band, in the heterochromatin of nuclei, and in the sarcoplasmic reticulum. The colocalization of HRG and skeletal muscle AMPD1 was also analyzed. A colabeling of HRG and AMPD1 was evident at sarcomeric, sarcoplasmic reticulum, and nuclear levels. The significance of these interesting and new results is discussed in this article.
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- 2019
11. Super-enhancer-driven metabolic reprogramming promotes cystogenesis in autosomal dominant polycystic kidney disease
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Zhiheng Liu, Bingxue Lan, Yongzhan Sun, Chaoran He, Hui Xiong, Xinyi Cao, Lirong Zhang, Yandong Song, Xu Zhu, Yi Lu, Zeyun Mi, Yupeng Chen, and Meijuan Geng
- Subjects
Male ,Endocrinology, Diabetes and Metabolism ,Autosomal dominant polycystic kidney disease ,Apoptosis ,Biology ,Phenylenediamines ,Kidney ,AMP Deaminase ,Mice ,Super-enhancer ,Physiology (medical) ,Internal Medicine ,medicine ,Animals ,Humans ,Enzyme Inhibitors ,Enhancer ,Gene ,Kinase ,Gene targeting ,Kidney metabolism ,Cell Biology ,medicine.disease ,Polycystic Kidney, Autosomal Dominant ,Adenosine Monophosphate ,Cyclin-Dependent Kinases ,Pyrimidines ,Gene Targeting ,Cancer research ,Female ,Cyclin-dependent kinase 7 ,Cyclin-Dependent Kinase-Activating Kinase - Abstract
Metabolic reprogramming is emerging as a key pathological contributor to the progression of autosomal dominant polycystic kidney disease (ADPKD), but the molecular mechanisms underlying dysregulated cellular metabolism in cystic cells remain elusive. Super-enhancers (SEs) are large clusters of transcriptional enhancers that drive robust expression of cell identity and disease genes. Here, we show that SEs undergo extensive remodelling during cystogenesis and that SE-associated transcripts are most enriched for metabolic processes in cystic cells. Inhibition of cyclin-dependent kinase 7 (CDK7), a transcriptional kinase required for assembly and maintenance of SEs, or AMP deaminase 3 (AMPD3), one of the SE-driven and CDK7-controlled metabolic target genes, delays cyst growth in ADPKD mouse models. In a cohort of people with ADPKD, CDK7 expression was frequently elevated, and its expression was correlated with AMPD3 expression and disease severity. Together, our findings elucidate a mechanism by which SE controls transcription of metabolic genes during cystogenesis, and identify SE-driven metabolic reprogramming as a promising therapeutic target for ADPKD treatment.
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- 2019
12. m6A RNA Degradation Products Are Catabolized by an Evolutionarily Conserved N6-Methyl-AMP Deaminase in Plant and Mammalian Cells
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Marco Herde, Claus-Peter Witte, Mounashree J. Urs, Mingjia Chen, Ismael Sánchez-González, and Monilola A. Olayioye
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0301 basic medicine ,Inosine monophosphate ,RNA Stability ,Arabidopsis ,Adenylate kinase ,Plant Science ,Biology ,In Brief ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,RNA polymerase ,Animals ,RNA, Messenger ,Research Articles ,Phylogeny ,Messenger RNA ,Arabidopsis Proteins ,Kinase ,fungi ,Eukaryota ,RNA ,AMP deaminase ,Cell Biology ,biology.organism_classification ,030104 developmental biology ,Biochemistry ,chemistry ,Transcriptome - Abstract
N(6)-methylated adenine (m(6)A) is the most frequent posttranscriptional modification in eukaryotic mRNA. Turnover of RNA generates N(6)-methylated AMP (N(6)-mAMP), which has an unclear metabolic fate. We show that Arabidopsis thaliana and human cells require an N(6)-mAMP deaminase (ADAL, renamed MAPDA) to catabolize N(6)-mAMP to inosine monophosphate in vivo by hydrolytically removing the aminomethyl group. A phylogenetic, structural, and biochemical analysis revealed that many fungi partially or fully lack MAPDA, which coincides with a minor role of N(6)A-RNA methylation in these organisms. MAPDA likely protects RNA from m(6)A misincorporation. This is required because eukaryotic RNA polymerase can use N(6)-mATP as a substrate. Upon abrogation of MAPDA, root growth is slightly reduced, and the N(6)-methyladenosine, N(6)-mAMP, and N(6)-mATP concentrations are increased in Arabidopsis. Although this will potentially lead to m(6)A misincorporation into RNA, we show that the frequency is too low to be reliably detected in vivo. Since N(6)-mAMP was severalfold more abundant than N(6)-mATP in MAPDA mutants, we speculate that additional molecular filters suppress the generation of N(6)-mATP. Enzyme kinetic data indicate that adenylate kinases represent such filters being highly selective for AMP versus N(6)-mAMP phosphorylation. We conclude that a multilayer molecular protection system is in place preventing N(6)-mAMP accumulation and salvage.
- Published
- 2018
13. Purine Nucleotides Metabolism and Signaling in Huntington’s Disease: Search for a Target for Novel Therapies
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Ewa M. Slominska, Talita Glaser, Marta Tomczyk, Henning Ulrich, and Ryszard T. Smolenski
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0301 basic medicine ,Purine ,congenital, hereditary, and neonatal diseases and abnormalities ,Huntingtin ,QH301-705.5 ,Review ,Biology ,Catalysis ,AMP Deaminase ,Inorganic Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Huntington's disease ,mental disorders ,medicine ,Animals ,Humans ,Nucleotide ,Biology (General) ,Enzyme Inhibitors ,Physical and Theoretical Chemistry ,Purine metabolism ,QD1-999 ,Purine Nucleotides ,Molecular Biology ,Spectroscopy ,chemistry.chemical_classification ,Huntingtin Protein ,DOENÇAS DEGENERATIVAS ,Catabolism ,Organic Chemistry ,Purinergic receptor ,purine metabolism ,General Medicine ,Purinergic signalling ,medicine.disease ,nervous system diseases ,Computer Science Applications ,Cell biology ,Chemistry ,Huntington Disease ,Neuroprotective Agents ,030104 developmental biology ,chemistry ,030217 neurology & neurosurgery ,purinergic signaling ,Huntington’s disease ,Signal Transduction - Abstract
Huntington’s disease (HD) is a multi-system disorder that is caused by expanded CAG repeats within the exon-1 of the huntingtin (HTT) gene that translate to the polyglutamine stretch in the HTT protein. HTT interacts with the proteins involved in gene transcription, endocytosis, and metabolism. HTT may also directly or indirectly affect purine metabolism and signaling. We aimed to review existing data and discuss the modulation of the purinergic system as a new therapeutic target in HD. Impaired intracellular nucleotide metabolism in the HD affected system (CNS, skeletal muscle and heart) may lead to extracellular accumulation of purine metabolites, its unusual catabolism, and modulation of purinergic signaling. The mechanisms of observed changes might be different in affected systems. Based on collected findings, compounds leading to purine and ATP pool reconstruction as well as purinergic receptor activity modulators, i.e., P2X7 receptor antagonists, may be applied for HD treatment.
- Published
- 2021
14. 5′-Adenosine monophosphate deaminase regulation in ground squirrels during hibernation
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Kenneth B. Storey and Khalil Abnous
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Hibernation ,medicine.medical_specialty ,Physiology ,Adenylate kinase ,030209 endocrinology & metabolism ,Biology ,01 natural sciences ,Biochemistry ,AMP Deaminase ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Animals ,Protein phosphorylation ,Energy charge ,Molecular Biology ,010405 organic chemistry ,Sciuridae ,Skeletal muscle ,AMP deaminase ,Torpor ,0104 chemical sciences ,Kinetics ,Endocrinology ,medicine.anatomical_structure ,Phosphorylation - Abstract
Hibernation is an important winter survival strategy for many small mammals. By sinking into a deep torpor where metabolic rate can be as low as 1–5% of the resting rate in euthermia, animals accrue huge energy savings that allow survival, typically without eating, for many months. Hibernating ground squirrels show a net reduction in the total adenylate pool of skeletal muscle during torpor, but the ATP/ADP ratio and adenylate energy charge remain stable. A key enzyme involved in managing adenylate pool size is 5′-adenosine monophosphate deaminase (AMPD). Assessing skeletal muscle AMPD from both Richardson's ground squirrels (Urocitellus richardsonii) (RGS) and 13-lined ground squirrels (Ictidomys tridecemlineatus) (TLGS), the present study shows that muscle AMPD of euthermic versus hibernating animals displays markedly different kinetic properties, differential responses to temperature and to effectors, and is regulated by reversible protein phosphorylation. AMPD activity decreased during hibernation in both TLGS and RGS skeletal muscle, by 70 and 84%, respectively. Stimulation of total protein phosphatases, total serine/threonine protein phosphatases, PP1, PP2B or PP2C, all reduced AMPD activity between 54 and 92% in extracts of euthermic RGS muscle. The same incubation did not change the activity of AMPD from muscle of hibernating animals. Oppositely, both euthermic and hibernating AMPD showed a strong increase in activity when incubated under conditions that promoted the enzyme phosphorylation by PKA, PKC or PKG. Overall, the data indicate that both low activity of AMPD and low affinity of the enzyme for AMP during torpor reduce the rate of adenylate degradation, the primary driver of these changes being covalent phosphorylation of AMPD.
- Published
- 2021
15. Effect of decaffeinated coffee on function and nucleotide metabolism in kidney
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Ryszard T. Smolenski, Ryszard Milczarek, Ewa M. Slominska, and Iwona Rybakowska
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0301 basic medicine ,medicine.medical_specialty ,Adenosine ,Kidney Cortex ,Clinical Biochemistry ,Decaffeinated coffee ,Purine nucleoside phosphorylase ,Renal function ,Coffee ,Article ,Ecto5′-nucleotidase ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Adenosine deaminase ,Internal medicine ,medicine ,Animals ,030212 general & internal medicine ,Molecular Biology ,Creatinine ,Kidney ,Kidney Medulla ,biology ,Nucleotides ,AMP deaminase ,Cell Biology ,General Medicine ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,chemistry ,biology.protein ,Caffeine ,medicine.drug - Abstract
Little is known about the effects of coffee that are not related to the presence of caffeine. The aim of the study was to analyse changes in kidney function and nucleotide metabolism related to high intake of decaffeinated coffee. Mice consumed decaffeinated coffee extract for two weeks. Activities of AMP deaminase, ecto5′-nucleotidase, adenosine deaminase, purine nucleoside phosphorylase were measured in kidney cortex and medulla by analysis of conversion of substrates into products using HPLC. Concentration of nucleotides in kidney cortex, kidney medulla and serum were estimated by HPLC. Activity of ecto5′-nucleotidase increased from 0.032 ± 0.006 to 0.049 ± 0.014 nmol/mg tissue/min in kidney cortex of mice administered high-dose decaffeinated coffee (HDC) together with increase in cortex adenosine concentration and decrease in plasma creatinine concentration. HDC leads to increased activity of ecto5′-nucleotidase in kidney cortex that translates to increase in concentration of adenosine. Surprisingly this caused improved kidney excretion function.
- Published
- 2017
16. The role of microorganisms in the degradation of adenosine triphosphate (ATP) in chill-stored common carp (Cyprinus carpio) fillets
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Sijia Song, Dapeng Li, Longteng Zhang, Zhiying Wang, Yongkang Luo, and Chunli Kong
- Subjects
Carps ,Microorganism ,Acid Phosphatase ,Food spoilage ,Population ,Colony Count, Microbial ,Cold storage ,Food Contamination ,Biology ,AMP Deaminase ,Analytical Chemistry ,Common carp ,chemistry.chemical_compound ,Adenosine Triphosphate ,0404 agricultural biotechnology ,Pseudomonas ,Animals ,education ,education.field_of_study ,04 agricultural and veterinary sciences ,General Medicine ,biology.organism_classification ,040401 food science ,Lactic acid ,Cold Temperature ,Lactobacillus ,Food Storage ,Seafood ,Biochemistry ,chemistry ,Food Microbiology ,Aeromonas ,Adenosine triphosphate ,Food Analysis ,Bacteria ,Food Science - Abstract
Biochemical and microbial changes after harvest strongly affect the final quality and shelf life of fish and fish products. In this study, the role of microbes in the degradation of adenosine triphosphate (ATP), and the origin of adenosine monophosphate deaminase (AMPD) and acid phosphatase (ACP) in common carp fillets during different stages of chilled storage (at 4°C) were investigated. The content of ATP, ADP, AMP, IMP, HxR, and Hx, the activity of AMPD and ACP, and the total count of viable, Aeromonas, Pseudomonas, H2S-producing bacteria, and lactic acid bacteria were examined. Results indicated that the population of microbial communities in control samples increased with storage time, and Pseudomonas peaked on the 10th day of storage. Changes in AMPD activity were less related to the abundance of microbes during the entire storage period. However, ACP was derived from both fish muscle and microbial secretion during the middle and late stages of storage. Degradation of ATP to IMP was not affected by spoilage bacteria, but the hydrolysis of IMP, and the transformation of HxR to Hx was affected considerably by the spoilage bacteria.
- Published
- 2017
17. Metabolic effects of combined introduction of adrenalin and blocker of methanoprolol beta-adrenophyleters
- Author
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A T Tapbergenov, Elina Hahn, S O Tapbergenov, and B S Sovetov
- Subjects
medicine.medical_specialty ,Epinephrine ,Adenosine Deaminase ,Glutathione reductase ,medicine.disease_cause ,Antioxidants ,General Biochemistry, Genetics and Molecular Biology ,AMP Deaminase ,5'-nucleotidase ,Adenosine deaminase ,Malondialdehyde ,Internal medicine ,Animals, Outbred Strains ,medicine ,Animals ,Sympathomimetics ,5'-Nucleotidase ,Metoprolol ,chemistry.chemical_classification ,Glutathione Peroxidase ,biology ,Chemistry ,Glutathione peroxidase ,Heart ,AMP deaminase ,General Medicine ,Catalase ,Adrenergic beta-1 Receptor Antagonists ,Rats ,Drug Combinations ,Glutathione Reductase ,Endocrinology ,Liver ,biology.protein ,Injections, Intraperitoneal ,Oxidative stress ,medicine.drug - Abstract
The effect of combined administration of adrenaline (0.4 mg/kg, i.p.) and b1-blocker metoprolol (25 mg/kg) on the activity of glutathione peroxidase (GPO), glutathione reductase (GR), catalase, adenosine deaminase (AD), AMP deaminase (AMPD), 5¢-nucleotidase (5¢N), on the level ofmalonic dialdehyde (MDA) and conjugated dienes (CD) was investigated. In blood adrenaline administration to animals caused an increase in the activity of AMPD, AD, 5¢N and GPO, and the increase the level of CD in the blood increases. Metoprolol caused a more pronounced increase in the activity of blood AMPD, AD, 5'N and the amount of CD. In contrast to adrenaline, metoprolol decreased the MDA level of, and decreased the activity of GPO and catalase. Combined administration of metoprolol and adrenaline to animals was accompanied by an increase in the activity of AD, AMPD, 5¢N, a decrease in the activity of GR, GPO, catalase, and a decrease in MDA in the blood. In the heart, adrenaline injection was accompanied by an increase in the MDA level, a decrease in 5¢N activity and an increase in the ratio of the activities of the enzymes AD+AMPD/5¢N. Metoprolol injection reduced MDA and CD levels and the activity of GR and GPO. The combined administration of metoprolol and adrenaline in the heart was accompanied by activation of AD, AMPD and 5¢N, and a decrease in the amount of MDA and CD, and a decrease in the activity of GR, GPO, and catalase. In the liver adrenaline caused an increase in MDA and DC levels, activation of catalase, AD, AMPD, and 5¢N. Metoprolol caused a decrease in MDA and CD levels and activity of catalase and GPO, an increase in the activity of AD and AMPD in the liver. Combined administration of adrenaline and metoprolol reduced manifestations of the heart and liver oxidative stress response as compared with administration of adrenaline alone.Izucheno vliianie sochetannogo vvedeniia adrenalina zhivotnym v doze 0,4 mg na 100 g massy, b1-blokatora metoprolola v doze 25 mg/kg na aktivnost' glutationperoksidazy (GPO), glutationreduktazy (GR), katalazy, adenozindezaminazy (AD), AMR-dezaminazy (AMPD), 5¢-nukleotidazy (5¢N), a takzhe na uroven' malonovogo dial'degida (MDA) i dienovykh kon'iugatov (DK). Vvedenie adrenalina privodilo k povysheniiu aktivnosti AMPD, AD, 5¢H i GPO, uvelichivalo uroven' DK v krovi. Metoprolol vyzyval bolee sil'noe uvelichenie aktivnosti AMPD, AD, 5¢H i kolichestva DK. V otlichie ot adrenalina, metoprolol snizhal uroven' MDA, aktivnost' GPO i katalazy. Sochetannoe vvedenie metoprolola i adrenalina vyzyvalo povyshenie aktivnosti AD, AMPD i 5¢H, snizhenie urovnia MDA i aktivnosti GR, GPO i katalazy v krovi. V serdtse vvedenie adrenalina privodilo k uvelicheniiu urovnia MDA, snizheniiu aktivnosti 5¢H i uvelicheniiu sootnosheniia aktivnosteĭ fermentov AD+AMPD/5¢N. Vvedenie metoprolola snizhalo uroven' MDA i DK v serdtse, aktivnost' GR i GPO, uvelichivalo sootnoshenie aktivnosti fermentov AD+AMPD/5'N. Sochetannoe vvedenie metoprolola i adrenalina vyzyvalo v serdtse aktivatsiiu AD, AMPD i 5¢H, snizhenie kolichestva MDA i DK. A takzhe snizhenie aktivnosti GR, GPO i katalazy. Pri vvedenie adrenalina otmecheno uvelichenie urovnia MDA i DK, aktivatsiia katalazy i fermentov metabolizma purinov AD, AMPD i 5¢H v pecheni. Éti dannye svidetel'stvuiut o tom, chto i v pecheni zhivotnykh, kak i v serdtse, pri vvedenii adrenalina proiskhodiat sdvigi, priblizhennye k sostoianiiu okislitel'nogo stressa. Pri vvedenii zhivotnym metoprolola v pecheni obnaruzheny snizhenie urovnia MDA i DK, i aktivnosti katalazy i GPO i povyshenie aktivnosti AD i AMPD. Pri sochetannom vvedenii adrenalina i metoprolola v pecheni, kak i v serdtse, otmecheno umen'shenie kolichestva MDA i DK, snizhenie aktivnosti GPO i katalazy, uvelichenie sootnosheniia aktivnosti fermentov AD+AMPD/5¢N.
- Published
- 2017
18. Dissociation of muscle insulin sensitivity from exercise endurance in mice by HDAC3 depletion
- Author
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Sisi Zhang, Mitchell A. Lazar, Wenjun Zhou, Henrik Molina, Yuxiang Zhang, Jennifer Jager, Zheng Sun, Manashree Damle, Bin Fang, Dan Feng, Guo-Lian Ding, Sungguan Hong, Brian D. Dill, Emanuele Loro, Tejvir S. Khurana, Qingwei Chu, Wenyun Lu, and Joshua D. Rabinowitz
- Subjects
Proteomics ,0301 basic medicine ,medicine.medical_specialty ,Blotting, Western ,Circadian clock ,Physical exercise ,Biology ,Real-Time Polymerase Chain Reaction ,Histone Deacetylases ,General Biochemistry, Genetics and Molecular Biology ,AMP Deaminase ,Epigenesis, Genetic ,Mice ,03 medical and health sciences ,Insulin resistance ,Lipid oxidation ,Physical Conditioning, Animal ,Internal medicine ,medicine ,Animals ,Muscle Strength ,Muscle, Skeletal ,Muscle fatigue ,Catabolism ,Skeletal muscle ,General Medicine ,Glucose clamp technique ,Lipid Metabolism ,medicine.disease ,Circadian Rhythm ,Histone Code ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Diabetes Mellitus, Type 2 ,Gene Knockdown Techniques ,Muscle Fatigue ,Body Composition ,Glucose Clamp Technique ,Physical Endurance ,Insulin Resistance ,Energy Metabolism ,Amino Acids, Branched-Chain - Abstract
Type 2 diabetes and insulin resistance are associated with reduced glucose utilization in the muscle and poor exercise performance. Here we find that depletion of the epigenome modifier histone deacetylase 3 (HDAC3) specifically in skeletal muscle causes severe systemic insulin resistance in mice but markedly enhances endurance and resistance to muscle fatigue, despite reducing muscle force. This seemingly paradoxical phenotype is due to lower glucose utilization and greater lipid oxidation in HDAC3-depleted muscles, a fuel switch caused by the activation of anaplerotic reactions driven by AMP deaminase 3 (Ampd3) and catabolism of branched-chain amino acids. These findings highlight the pivotal role of amino acid catabolism in muscle fatigue and type 2 diabetes pathogenesis. Further, as genome occupancy of HDAC3 in skeletal muscle is controlled by the circadian clock, these results delineate an epigenomic regulatory mechanism through which the circadian clock governs skeletal muscle bioenergetics. These findings suggest that physical exercise at certain times of the day or pharmacological targeting of HDAC3 could potentially be harnessed to alter systemic fuel metabolism and exercise performance.
- Published
- 2016
19. Adenosine monophosphate deaminase 3 activation shortens erythrocyte half-life and provides malaria resistance in mice
- Author
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Denis C. Bauer, Elinor Hortle, Dedreia Tull, Ian A. Cockburn, Shelley Lampkin, Lora M. Jensen, Simon J. Foote, Seong Beom Ahn, Gaetan Burgio, Brendan J. McMorran, Malcolm J. McConville, and Brunda Nijagal
- Subjects
Male ,0301 basic medicine ,Senescence ,medicine.medical_specialty ,Erythrocytes ,Thalassemia ,Immunology ,Biology ,Biochemistry ,AMP Deaminase ,Plasmodium chabaudi ,Mice ,03 medical and health sciences ,Red Cells, Iron, and Erythropoiesis ,0302 clinical medicine ,Internal medicine ,parasitic diseases ,medicine ,Animals ,Erythropoiesis ,Cellular Senescence ,1102 Cardiorespiratory Medicine and Haematology, 1103 Clinical Sciences, 1114 Paediatrics and Reproductive Medicine ,hemic and immune systems ,AMP deaminase ,Cell Biology ,Hematology ,biology.organism_classification ,medicine.disease ,Adenosine ,Immunity, Innate ,Sickle cell anemia ,Malaria ,Red blood cell ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Ethylnitrosourea ,030220 oncology & carcinogenesis ,Mutation ,Half-Life ,circulatory and respiratory physiology ,medicine.drug - Abstract
The factors that determine red blood cell (RBC) lifespan and the rate of RBC aging have not been fully elucidated. In several genetic conditions, including sickle cell disease, thalassemia, and G6PD deficiency, erythrocyte lifespan is significantly shortened. Many of these diseases are also associated with protection from severe malaria, suggesting a role for accelerated RBC senescence and clearance in malaria resistance. Here, we report a novel, N-ethyl-N-nitrosourea-induced mutation that causes a gain of function in adenosine 5'-monophosphate deaminase (AMPD3). Mice carrying the mutation exhibit rapid RBC turnover, with increased erythropoiesis, dramatically shortened RBC lifespan, and signs of increased RBC senescence/eryptosis, suggesting a key role for AMPD3 in determining RBC half-life. Mice were also found to be resistant to infection with the rodent malaria Plasmodium chabaudi. We propose that resistance to P. chabaudi is mediated by increased RBC turnover and higher rates of erythropoiesis during infection.
- Published
- 2016
20. Safety evaluation of AMP deaminase from Aspergillus oryzae
- Author
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Ashley Roberts, Maya Ueda, Mai Sugi, Barry Lynch, Nobuo Okado, Fukutaro Mizuhashi, and Trung D. Vo
- Subjects
Male ,Salmonella typhimurium ,Inosine monophosphate ,Dose ,Aspergillus oryzae ,Administration, Oral ,Cadherin Related Proteins ,Toxicology ,medicine.disease_cause ,Chromosome aberration ,AMP Deaminase ,Ames test ,Cricetinae ,medicine ,Animals ,Food science ,Protein Precursors ,Cells, Cultured ,biology ,Mutagenicity Tests ,Chemistry ,AMP deaminase ,General Medicine ,Fibroblasts ,Cadherins ,biology.organism_classification ,Rats ,Biochemistry ,Female ,Fermentation ,Genotoxicity ,Food Science - Abstract
Adenosine-5'-monophosphate (AMP) deaminase is an enzyme used to increase concentrations of 5'-inosine monophosphate in certain foods and beverages for flavoring purposes. One commercial source of this enzyme is Aspergillus oryzae, a filamentous fungus with a history of safe use in Asia as a fermentation organism used in the production of miso sauce and sake liquors. Noting the use of the enzyme in food intended for human consumption and potential presence at trace levels in finished goods, a series of safety studies including an in vitro Ames test and chromosome aberration assay with Chinese hamster lung fibroblasts were conducted along with a 90-day oral toxicity study in rats. AMP deaminase showed no evidence of genotoxicity in the in vitro tests. Following gavage administration of Sprague-Dawley rats at dosages of 19.8, 198.4, or 1984 mg total organic solids (TOS)/kg body weight (bw)/day for 90 days, no adverse effects on body weight gain, food consumption, hematology, clinical chemistry, urinalysis, ophthalmological and histopathological examinations were observed. The no-observed-adverse-effect level was considered to be 1984 mg TOS/kg bw/day, the highest dose tested. Results of the genotoxicity studies and subchronic rat study support the safe use of AMP deaminase produced from A. oryzae in food production.
- Published
- 2015
21. Identification of Two Novel Single Nucleotide Polymorphisms in the Promoter Region of the Pig AMP Deaminase 1 Gene Associated with Carcass Traits
- Author
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Zhe Chao, Linjie Wang, and Yan Wang
- Subjects
Male ,Meat ,Swine ,Energy metabolism ,Gene Expression ,Single-nucleotide polymorphism ,Biology ,Polymorphism, Single Nucleotide ,AMP Deaminase ,Quantitative Trait, Heritable ,Genetics ,SNP ,Animals ,Body Fat Distribution ,Muscle, Skeletal ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Alleles ,Purine nucleotide interconversion ,Base Sequence ,Body Weight ,Promoter ,AMP deaminase ,Cell Biology ,General Medicine ,Phenotype ,Adipose Tissue ,Haplotypes ,Genetic Loci ,Identification (biology) ,Female - Abstract
The AMP deaminase 1 (AMPD1) gene plays an important role in purine nucleotide interconversion and energy metabolism. In this study, two novel single nucleotide polymorphisms (SNPs) (g.-626 G A and g.-566 AG) were detected in the proximal promoter region of the AMPD1 gene. The Chinese indigenous pig breeds (Meishan and Tongcheng) had higher frequencies of the G and A alleles than Western meat-type breeds (Landrace and Large White) at the g.-626 G A and g.-566 AG loci. The transcriptional activity of the AMPD1 promoter carrying the haplotype H1 (A
- Published
- 2018
22. Functional genomics identifies AMPD2 as a new prognostic marker for undifferentiated pleomorphic sarcoma
- Author
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Martin F, Orth, Julia S, Gerke, Thomas, Knösel, Annelore, Altendorf-Hofmann, Julian, Musa, Rebeca, Alba-Rubio, Stefanie, Stein, Tilman L B, Hölting, Florencia, Cidre-Aranaz, Laura, Romero-Pérez, Marlene, Dallmayer, Michaela C, Baldauf, Aruna, Marchetto, Giuseppina, Sannino, Maximilian M L, Knott, Fabienne, Wehweck, Shunya, Ohmura, Jing, Li, Michiyuki, Hakozaki, Thomas, Kirchner, Thomas, Dandekar, Elke, Butt, and Thomas G P, Grünewald
- Subjects
Adult ,Aged, 80 and over ,Mice, Knockout ,Gene Expression Profiling ,Genomics ,Histiocytoma, Malignant Fibrous ,Kaplan-Meier Estimate ,Mice, SCID ,Middle Aged ,Prognosis ,Xenograft Model Antitumor Assays ,AMP Deaminase ,Gene Expression Regulation, Neoplastic ,Young Adult ,RNAi Therapeutics ,Mice, Inbred NOD ,Cell Line, Tumor ,Biomarkers, Tumor ,Animals ,Humans ,Aged - Abstract
Soft-tissue sarcomas are rare, heterogeneous, and often aggressive mesenchymal cancers. Many of them are associated with poor outcome, partially because biomarkers that can identify high-risk patients are lacking. Studies on sarcomas are often limited by small sample-sizes rendering the identification of biomarkers difficult when focusing on individual cohorts. However, the increasing number of publicly available 'omics' data opens inroads to overcome this obstacle. Here, we combine transcriptome analyses, immunohistochemistry, and functional assays to show that high adenosine monophosphate deaminase 2 (AMPD2) is a robust prognostic biomarker for worse outcome in undifferentiated pleomorphic sarcoma (UPS). Gene expression and survival data for UPS from two independent studies were subjected to survival association-testing. Genes, whose high expression was significantly correlated with worse outcome in both cohorts, were considered as biomarker candidates. The best candidate, AMPD2, was validated in a tissue microarray. Analysis of DNA copy-number data and matched transcriptomes indicated that high AMPD2 expression is significantly correlated with gains at the AMPD2 locus. Gene set enrichment analyses of AMPD2 co-expressed genes in both transcriptome datasets suggested that AMPD2-high UPS are enriched in tumorigenic signatures. Consistently, knockdown of AMPD2 by RNA interference in an UPS cell line inhibited proliferation in vitro and tumorigenicity in vivo. Collectively, we provide evidence that AMPD2 may serve as a biomarker for outcome prediction in UPS. Our study exemplifies how the integration of 'omics' data, immunohistochemistry, and functional experiments can identify novel biomarkers even in a rare sarcoma, which may serve as a blueprint for biomarker identification for other rare cancers.
- Published
- 2018
23. Growth and Metabolic Response of Chinese Perch to Different Dietary Protein-to-Energy Ratios in Artificial Diets
- Author
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Xu-Fang Liang, Shan He, Yulan Kuang, Muhammad Shoaib Alam, Farman Ullah Dawar, Seyed Hossein Hoseinifar, and Liwei Liu
- Subjects
AMPK ,0301 basic medicine ,Pro-Opiomelanocortin ,Protein efficiency ratio ,protein synthesis ,nitrogen metabolism ,AMP Deaminase ,chemistry.chemical_compound ,Predatory fish ,AMP-Activated Protein Kinase Kinases ,Glutamate Dehydrogenase ,growth and metabolism ,Spectroscopy ,Perch ,biology ,TOR Serine-Threonine Kinases ,Alanine Transaminase ,04 agricultural and veterinary sciences ,General Medicine ,Computer Science Applications ,Animal Nutritional Physiological Phenomena ,Nutrition physiology ,Dietary Proteins ,medicine.symptom ,Fish Proteins ,Fisheries ,Chinese perch ,mTOR pathways ,Feed conversion ratio ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Animal science ,medicine ,Animals ,Aspartate Aminotransferases ,Physical and Theoretical Chemistry ,energy homeostasis ,Molecular Biology ,body composition ,Triglyceride ,Organic Chemistry ,biology.organism_classification ,Diet ,030104 developmental biology ,chemistry ,Blood chemistry ,Perches ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,protein/energy ratio ,Energy Metabolism ,Protein Kinases ,Weight gain - Abstract
The effect of dietary nutrients on novel farm species has always garnered wide research and economic interest. Chinese perch, an economically important carnivorous fish, accepts an artificial diet after taming, so it is essential to evaluate and optimize the nutritional and metabolic demands of this species. However, little is known about the effect of an artificial diet on the growth and metabolism of Chinese perch. Therefore, the present study evaluated the growth and metabolic responses of Chinese perch to experimental diets with different dietary protein/energy (P/E) ratios. Five isoenergetic diets (18 kJ/g) with graded levels of P/E ratios of 30.58, 33.22, 35.90, 38.6, and 41.35 mg/kJ (named A, B, C, D, and E) were formulated. A total of 225 Chinese perch (64.89 ±, 0.28 g) were divided into five groups (triplicate tanks for each group), distributed into 15 (350 L) fiberglass tanks, and fed twice a day at 4% of fish wet body weight with the respective P/E ratio diets for 10 weeks. Compared with the other groups, Chinese perch in Group C showed significantly improved growth performance, weight gain (WG), specific growth rate (SGR), viscerosomatic index (VSI), hepatosomatic index (HSI), intraperitoneal fat (IPF), feed utilization, feed intake (FI), feed conversion ratio (FCR), protein efficiency ratio (PER), protein retention efficiency (PRE), energy retention efficiency (ERE), and feed efficiency (FE) as well as whole-body, muscle, and liver composition. Chinese perch in Group A, on the other hand, had the lowest growth performance, feed utilization, and body composition compared with the other groups. The activities of nitrogen metabolism-related enzymes (alanine aminotransferase (ALT), aspartate aminotransferase (AST) glutamate dehydrogenase (GDH), and adenosine 5&prime, monophosphate deaminase (AMPD)) as well as the mRNA expression of the GDH and AMPD genes were significantly lower than those in the other groups. Similarly, the expression of NPY and AgRp were significantly higher in Group C compared with the other groups. However, the gene expression of CART and POMC was not affected by the dietary P/E ratios. In Group A, the expression of mTOR, S6K, and 4EBP1 was significantly lower and that of AMPK, LKB1, and eEF2 was significantly higher when compared with the other groups. Biochemical analysis of blood showed that ALT, AST, total protein (TP), alkaline phosphatase (ALP), glucose (GLU), blood urea nitrogen (BUN), and triglyceride (TG) levels were also affected by the dietary P/E ratio. From our results, we concluded that Chinese perch growth performance and nutrient metabolism were significantly affected by the P/E ratio of the artificial diet. Second-order polynomial regression analysis revealed that Chinese perch growth performance was optimal at a P/E ratio of 37.98 in the artificial diet.
- Published
- 2019
24. Role of the HPRG Component of Striated Muscle AMP Deaminase in the Stability and Cellular Behaviour of the Enzyme
- Author
-
Antonio Raggi and Francesca Ronca
- Subjects
Gene isoform ,Protein subunit ,lcsh:QR1-502 ,Review ,Biochemistry ,lcsh:Microbiology ,AMP Deaminase ,Enzyme Stability ,medicine ,Animals ,Humans ,Muscle, Skeletal ,Molecular Biology ,Actin ,chemistry.chemical_classification ,biology ,Skeletal muscle ,Proteins ,Troponin T (TnT) ,AMP deaminase ,Amp deaminase (AMPD) ,Histidine-proline-rich glycoprotein (HPRG) ,Striated muscle ,Troponin T (TNT) ,Alternative Splicing ,Isoenzymes ,Skeletal ,AMP deaminase (AMPD) ,Cell biology ,histidine-proline-rich glycoprotein (HPRG) ,striated muscle ,medicine.anatomical_structure ,Enzyme ,chemistry ,Chaperone (protein) ,biology.protein ,Muscle ,Glycoprotein - Abstract
Multiple muscle-specific isoforms of the Zn2+ metalloenzyme AMP deaminase (AMPD) have been identified based on their biochemical and genetic differences. Our previous observations suggested that the metal binding protein histidine-proline-rich glycoprotein (HPRG) participates in the assembly and maintenance of skeletal muscle AMP deaminase (AMPD1) by acting as a zinc chaperone. The evidence of a role of millimolar-strength phosphate in stabilizing the AMPD-HPRG complex of both AMPD1 and cardiac AMP deaminase (AMPD3) is suggestive of a physiological mutual dependence between the two subunit components with regard to the stability of the two isoforms of striated muscle AMPD. The observed influence of the HPRG content on the catalytic behavior of the two enzymes further strengthens this hypothesis. Based on the preferential localization of HPRG at the sarcomeric I-band and on the presence of a Zn2+ binding motif in the N-terminal regions of fast TnT and of the AMPD1 catalytic subunit, we advance the hypothesis that the Zn binding properties of HPRG could promote the association of AMPD1 to the thin filament.
- Published
- 2018
25. Effect of quercetin on E-NTPDase/E-ADA activities and cytokine secretion of complete Freund adjuvant-induced arthritic rats
- Author
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Alessandra G. Manzoni, Nara Maria Beck Martins, Lívia G. Castilhos, Juliana Sorraila de Oliveira, Daniela B.R. Leal, Lara Vargas Becker, Emerson André Casali, Stephen A. Adefegha, Fátima Husein Abdalla, Ana Paula Visintainer Coelho, Leonardo Lanes da Silveira, Karine Lanes da Silveira, and Renata da Silva Pereira Saccol
- Subjects
0301 basic medicine ,Clinical Biochemistry ,Flavonoid ,Freund's Adjuvant ,Arthritis ,Pharmacology ,Biochemistry ,AMP Deaminase ,Arthritis, Rheumatoid ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Immune system ,Medicine ,Animals ,Edema ,heterocyclic compounds ,Enzyme Inhibitors ,Rats, Wistar ,chemistry.chemical_classification ,Adenosine Triphosphatases ,biology ,business.industry ,Anti-Inflammatory Agents, Non-Steroidal ,Cell Biology ,General Medicine ,medicine.disease ,Adenosine ,Rats ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Rheumatoid arthritis ,Myeloperoxidase ,biology.protein ,Cytokines ,Cytokine secretion ,Female ,Quercetin ,business ,medicine.drug - Abstract
The effect of quercetin was assessed in rats induced with complete Freund adjuvant (CFA). Arthritis scores, paw oedema, latency, activities of myeloperoxidase (MPO), ectonucleoside triphosphate diphosphohydrolase (E-NTPDase), and ectoadenosine deaminase (E-ADA) in lymphocytes were determined. Furthermore, nucleotide and nucleoside levels as well as the secretion of pro- and anti-inflammatory cytokines were evaluated. Animals were treated with saline and quercetin in doses of 5, 25, and 50 mg/kg for 45 days. The result revealed that quercetin (50 mg/kg) reduced arthritis score and paw oedema, and increased the latency in the thermal hyperalgesia test. Histopathological analysis showed that all the doses of quercetin reduced infiltration of inflammatory cells. MPO activity was increased in the arthritis group; however, quercetin reduced this activity. E-NTPDase activity was increased in lymphocytes of arthritis rats, and treatment with quercetin reversed this increase. However, E-ADA activity was reduced in the arthritis group, and treatment with quercetin modulated the activity of this enzyme in arthritis rat groups. Serum adenosine levels were increased in arthritis, and the levels were lowered with quercetin treatment. Quercetin treatment in arthritis groups decreased the elevated levels of cytokines in the arthritis control group. Thus, quercetin demonstrated an anti-inflammatory effect, and this flavonoid may be a promising natural compound for the treatment of arthritis. SIGNIFICANCE OF THE STUDY: Quercetin may represent a potential therapeutic compound in the treatment of rheumatoid arthritis. Findings from this study indicate that quercetin suppresses swelling and attenuates the underlying inflammatory responses. This is the first report where quercetin was shown to modulate the immune response to arthritis via attenuation of the purinergic system (E-NTPDase and E-ADA activities) and the levels of IFN-gamma and IL-4. Thus, this work is relevant to basic research and may be translated into clinical practice.
- Published
- 2018
26. Uric Acid as a Cause of the Metabolic Syndrome
- Author
-
Christopher, King, Miguel A, Lanaspa, Thomas, Jensen, Dean R, Tolan, L Gabriela, Sánchez-Lozada, and Richard J, Johnson
- Subjects
Metabolic Syndrome ,Dietary Sugars ,Fructose ,Hyperuricemia ,AMP Deaminase ,Mitochondria ,Uric Acid ,Fatty Liver ,Oxidative Stress ,Hypertension ,Diabetes Mellitus ,Animals ,Humans ,Insulin Resistance - Abstract
Hyperuricemia is common in subjects with obesity, metabolic syndrome, and type 2 diabetes. For many years, hyperuricemia was attributed to the effects of insulin resistance to reduce urinary excretion of uric acid, and it was believed that uric acid may not have any causal role in the metabolic syndrome. However, in recent years, hyperuricemia has been found to independently predict the development of diabetes. Experimental studies have also shown that hyperuricemia may mediate insulin resistance, fatty liver, and dyslipidemia in both fructose-dependent and fructose-independent models of metabolic syndrome. The mechanism for uric acid-induced insulin resistance appears to be mediated by the development of mitochondrial oxidative stress and impairment of insulin-dependent stimulation of nitric oxide in endothelial cells. Pilot studies in humans have reported a potential benefit of lowering serum uric acid on insulin resistance. Large clinical trials are recommended. If uric acid is shown to be a mediator of incident type 2 diabetes in humans, then lowering serum uric acid would represent a simple and inexpensive way to help prevent the development of diabetes and to slow the epidemic.
- Published
- 2018
27. Translational regulation by miR-301b upregulates AMP deaminase in diabetic hearts
- Author
-
Koki Abe, Satoko Ishikawa, Atsushi Kuno, Hiromu Suzuki, Toshiyuki Yano, Takayuki Miki, Tatsuya Sato, Masaya Tanno, Tetsuji Miura, Yuki Tatekoshi, Takeshi Niinuma, Wataru Ohwada, and Hidemichi Kouzu
- Subjects
0301 basic medicine ,medicine.medical_specialty ,030204 cardiovascular system & hematology ,AMP Deaminase ,03 medical and health sciences ,0302 clinical medicine ,Adenosine Triphosphate ,Downregulation and upregulation ,Adenine nucleotide ,Internal medicine ,Translational regulation ,microRNA ,medicine ,Animals ,Humans ,Myocytes, Cardiac ,Molecular Biology ,Messenger RNA ,Chemistry ,Adenine ,Myocardium ,AMP deaminase ,Transfection ,Myocardial Contraction ,Rats ,Disease Models, Animal ,MicroRNAs ,030104 developmental biology ,Endocrinology ,Diabetes Mellitus, Type 2 ,Gene Expression Regulation ,Proteasome inhibitor ,Cardiology and Cardiovascular Medicine ,medicine.drug - Abstract
AMP deaminase (AMPD) plays a crucial role in adenine nucleotide metabolism. Recently we found that upregulated AMPD activity is associated with ATP depletion and contractile dysfunction under the condition of pressure overloading in the heart of a rat model of type 2 diabetes mellitus (T2DM), OLETF. Here we examined the mechanism of AMPD upregulation by T2DM. The protein level of 90-kDa full-length AMPD3 was increased in whole myocardial lysates by 55% in OLETF compared to those in LETO, a non-diabetic control. In contrast, the mRNA levels of AMPD3 in the myocardium were similar in OLETF and LETO. AMPD3 was comparably ubiquitinated in OLETF and LETO, and its degradation ex vivo was more sensitive to MG-132, a proteasome inhibitor, in OLETF than in LETO. MicroRNA array analysis revealed downregulation (>50%) of 57 microRNAs in OLETF compared to those in LETO, among which miR-301b was predicted to interact with the 3'UTR of AMPD3 mRNA. AMPD3 protein level was significantly increased by a miR-301b inhibitor and was decreased by a miR-301b mimetic in H9c2 cells. A luciferase reporter assay confirmed binding of miR-301b to the 3'UTR of AMPD3 mRNA. Transfection of neonatal rat cardiomyocytes with a miR-301b inhibitor increased 90-kDa AMPD3 and reduced ATP level. The results indicate that translational regulation by miR-301b mediates upregulated expression of cardiac AMPD3 protein in OLETF, which potentially reduces the adenine nucleotide pool at the time of increased work load. The miR-301b-AMPD3 axis may be a novel therapeutic target for intervening enegy metabolism in diabetic hearts.
- Published
- 2017
28. HIF-1α in the heart: Remodeling nucleotide metabolism
- Author
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Ying Li, Cherie E. Bond, Ping Chen, Minghua Chen, Gary L. Wright, Ralph V. Shohet, and Joe Wu
- Subjects
Time Factors ,Ischemia ,Gene Expression ,Adenylate kinase ,Mice, Transgenic ,Biology ,Article ,AMP Deaminase ,Mice ,chemistry.chemical_compound ,medicine ,Animals ,Energy charge ,Molecular Biology ,Hypoxanthine ,Nucleotides ,Myocardium ,Purine nucleotide cycle ,Heart ,AMP deaminase ,Hypoxia-Inducible Factor 1, alpha Subunit ,medicine.disease ,Molecular biology ,Adenosine ,chemistry ,Hypoxanthine-guanine phosphoribosyltransferase ,Models, Animal ,Cardiology and Cardiovascular Medicine ,medicine.drug - Abstract
These studies have examined the effect of hypoxia inducible factor 1α (HIF-1α) on nucleotide metabolism in the ischemic heart using a genetic mouse model with heart-specific and regulated expression of a stable form of HIF-1α. We find that AMP deaminase (AMPD), the entry point of the purine nucleotide cycle (PNC), is induced by HIF-1α at the level of mRNA, protein, and activity. AMP that accumulates during ischemia can be metabolized to adenosine by 5′-nucleotidase or to IMP by AMPD. Consistent with the finding of AMPD induction, adenosine accumulation during ischemia was much attenuated in HIF-1α-expressing hearts. Further investigation of nucleotide salvage enzymes found that hypoxanthine phosphoribosyl transferase (HPRT) is also upregulated in HIF-1α-expressing hearts. Treatment of hearts with an inhibitor of the PNC, hadacidin, hastens the fall of the adenylate energy charge during ischemia and the accumulation of AMP. The results provide new insight into the role of the PNC in the heart, especially as it relates to ischemia, and indicate that HIF-1α regulates nucleotide metabolism as a compensatory response to hypoxia.
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- 2015
29. Excessive degradation of adenine nucleotides by up-regulated AMP deaminase underlies afterload-induced diastolic dysfunction in the type 2 diabetic heart
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Makoto Ogasawara, Tatsuya Sato, Toshiyuki Yano, Toshiyuki Tobisawa, Hiromichi Murase, Hidemichi Kouzu, Atsushi Kuno, Masaya Tanno, Takayuki Miki, Tetsuji Miura, Satoko Ishikawa, Daisuke Sunaga, and Takahito Itoh
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medicine.medical_specialty ,Diastole ,Gene Expression ,AMP Deaminase ,Afterload ,Adenine nucleotide ,Internal medicine ,medicine ,Animals ,Metabolomics ,Protein Isoforms ,Ventricular Function ,Connectin ,Phosphorylation ,Molecular Biology ,Heart Failure, Diastolic ,Adenine Nucleotides ,Chemistry ,Myocardium ,Hemodynamics ,Diastolic heart failure ,AMP deaminase ,medicine.disease ,Myocardial Contraction ,Rats ,Disease Models, Animal ,Preload ,Endocrinology ,Diabetes Mellitus, Type 2 ,Heart failure ,Heart Function Tests ,Metabolome ,Aortic pressure ,Cardiology and Cardiovascular Medicine - Abstract
Type 2 diabetes mellitus (T2DM) is often complicated with diastolic heart failure, which decompensates under increased afterload. Focusing on cardiac metabolomes, we examined mechanisms by which T2DM augments ventricular diastolic stiffness in response to pressure overloading. Pressure-volume relationships (PVRs) and myocardial metabolomes were determined at baseline and during elevation of aortic pressure by phenylephrine infusion in a model of T2DM, OLETF, and its non-diabetic control, LETO. Pressure overloading augmented diastolic stiffness without change in systolic reserve in OLETF as indicated by a left-upward shift of end-diastolic PVR. In contrast, PVRs under cardioplegic arrest in buffer-perfused isolated hearts were similar in OLETF and LETO, indicating that extracellular matrix or titin remodeling does not contribute to the afterload-induced increase in stiffness of the beating ventricle of OLETF. Metabolome analyses revealed impaired glycolysis and facilitation of the pentose phosphate pathway in OLETF. Pressure overloading significantly reduced ATP and total adenine nucleotides by 34% and 40%, respectively, in OLETF but not in LETO, while NADH-to-NAD(+) ratios were similar in the two groups. The decline in ATP by pressure overloading in OLETF was associated with increased inosine 5-monophosphate and decreased adenosine levels, being consistent with the 2.5-times higher activity of cardiac AMP deaminase in OLETF. Tissue ATP level was negatively correlated with tau of LV pressure and LVEDP. These results suggest that ATP depletion due to excessive degradation of adenine nucleotides by up-regulated AMP deaminase underlies ventricular stiffening during acute pressure overloading in T2DM hearts.
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- 2015
30. DNA sequence polymorphism within the bovine adenosine monophosphate deaminase 1 (AMPD1) is associated with production traits in Chinese cattle
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Juqiang Wang, F.-Y. Chen, C.-B. Wei, H. Niu, and K. Li
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Meat ,Genotype ,Breeding ,Biology ,Beef cattle ,Quantitative trait locus ,AMP Deaminase ,law.invention ,Quantitative Trait, Heritable ,law ,Genetics ,Animals ,Allele ,Molecular Biology ,Gene ,Genetic Association Studies ,Polymerase chain reaction ,Sequence Deletion ,Body Weight ,AMP deaminase ,Heterozygote advantage ,Sequence Analysis, DNA ,General Medicine ,Molecular biology ,Phenotype ,Cattle - Abstract
The objectives of the present study were to detect an 18-bp deletion mutation in the bovine adenosine monophosphate deaminase 1 (AMPD1) gene and analyze its effect on growth traits in 2 Chinese cattle breeds using DNA sequencing and agarose electrophoresis. The five 19-bp polymerase chain reaction products of the AMPD1 gene exhibited 3 genotypes and 2 alleles: WW: homozygote genotype (wild-type); DD: homozygote genotype (mutant-type); WD: heterozygote genotype. Frequencies of the W allele varied from 66.15-70.35%. The associations between the 18-bp deletion mutation in the AMPD1 gene with production traits in 226 Jia-Xian red cattle was analyzed. The animals with genotype WW showed significantly higher heart girth and body weight than those with genotypes WD and DD at 24 months (P < 0.01). Our results indicate that the deletion mutation in the AMPD1 gene is associated with production traits, and may be used for marker-assisted selection in beef cattle breeding programs.
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- 2015
31. Effects of Pharmacological AMP Deaminase Inhibition and Ampd1 Deletion on Nucleotide Levels and AMPK Activation in Contracting Skeletal Muscle
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Louis Hue, Nadège Zanou, Mark H. Rider, Samanta Kviklyte, Lars Löfgren, Catheline Plaideau, Philippe Gailly, Yu-Chiang Lai, Didier Vertommen, Mohammad Bohlooly-Y, Harriet Andersén, and Stefan Hallén
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Male ,medicine.medical_specialty ,Clinical Biochemistry ,Stimulation ,AMP-Activated Protein Kinases ,In Vitro Techniques ,Biology ,Biochemistry ,AMP Deaminase ,Mice ,Adenosine Triphosphate ,Adenine nucleotide ,Internal medicine ,Drug Discovery ,medicine ,Animals ,Enzyme Inhibitors ,Rats, Wistar ,Muscle, Skeletal ,Protein kinase A ,Purine Nucleotides ,Molecular Biology ,Mice, Knockout ,Pharmacology ,Glucose transporter ,Skeletal muscle ,AMPK ,AMP deaminase ,General Medicine ,Adenosine Monophosphate ,Electric Stimulation ,Rats ,Mice, Inbred C57BL ,Glucose ,medicine.anatomical_structure ,Endocrinology ,Phosphorylation ,Molecular Medicine ,Acetyl-CoA Carboxylase ,Muscle Contraction - Abstract
SummaryAMP-activated protein kinase (AMPK) plays a central role in regulating metabolism and energy homeostasis. It achieves its function by sensing fluctuations in the AMP:ATP ratio. AMP deaminase (AMPD) converts AMP into IMP, and the AMPD1 isoenzyme is expressed in skeletal muscles. Here, effects of pharmacological inhibition and genetic deletion of AMPD were examined in contracting skeletal muscles. Pharmacological AMPD inhibition potentiated rises in AMP, AMP:ATP ratio, AMPK Thr172, and acetyl-CoA carboxylase (ACC) Ser218 phosphorylation induced by electrical stimulation, without affecting glucose transport. In incubated extensor digitorum longus and soleus muscles from Ampd1 knockout mice, increases in AMP levels and AMP:ATP ratio by electrical stimulation were potentiated considerably compared with muscles from wild-type mice, whereas enhanced AMPK activation was moderate and only observed in soleus, suggesting control by factors other than changes in adenine nucleotides. AMPD inhibitors could be useful tools for enhancing AMPK activation in cells and tissues during ATP-depletion.
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- 2014
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32. Inhibition of AMP Deaminase Activity Does Not Improve Glucose Control in Rodent Models of Insulin Resistance or Diabetes
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Lars Löfgren, Tomas Drmota, Kristina Wallenius, Stefan Hallén, Jianming Liu, Nidhal Selmi, Mattias Rohman, Martin Bauer, Bo Lindmark, Therese Admyre, Judith Hartleib-Geschwindner, Mikael Bjursell, Lena Amrot-Fors, and Maria Andersson
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Blood Glucose ,medicine.medical_specialty ,AMP deaminase activity ,Clinical Biochemistry ,Mice, Obese ,Biology ,Diet, High-Fat ,Biochemistry ,AMP Deaminase ,Diabetes Mellitus, Experimental ,Rats, Sprague-Dawley ,Small Molecule Libraries ,Mice ,Insulin resistance ,In vivo ,Internal medicine ,Drug Discovery ,medicine ,Animals ,Insulin ,Obesity ,Enzyme Inhibitors ,Protein kinase A ,Molecular Biology ,Cells, Cultured ,Mice, Knockout ,Pharmacology ,AMPK ,AMP deaminase ,General Medicine ,medicine.disease ,Adenosine ,Recombinant Proteins ,Rats ,Mice, Inbred C57BL ,Disease Models, Animal ,Endocrinology ,Hepatocytes ,Molecular Medicine ,Female ,Insulin Resistance ,Ex vivo ,Protein Binding ,medicine.drug - Abstract
SummaryInhibition of AMP deaminase (AMPD) holds the potential to elevate intracellular adenosine and AMP levels and, therefore, to augment adenosine signaling and activation of AMP-activated protein kinase (AMPK). To test the latter hypothesis, novel AMPD pan inhibitors were synthesized and explored using a panel of in vitro, ex vivo, and in vivo models focusing on confirming AMPD inhibitory potency and the potential of AMPD inhibition to improve glucose control in vivo. Repeated dosing of selected inhibitors did not improve glucose control in insulin-resistant or diabetic rodent disease models. Mice with genetic deletion of the muscle-specific isoform Ampd1 did not show any favorable metabolic phenotype despite being challenged with high-fat diet feeding. Therefore, these results do not support the development of AMPD inhibitors for the treatment of type 2 diabetes.
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- 2014
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33. New insights on the regulation of the adenine nucleotide pool of erythrocytes in mouse models
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Han Shawn Ling, Zhaoyang Zhao, William G. O’Brien, and Cheng Chi Lee
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0301 basic medicine ,Thin-Layer Chromatography ,Adenosine ,Erythrocytes ,Time Factors ,Physiology ,Glycobiology ,lcsh:Medicine ,Biochemistry ,5'-nucleotidase ,AMP Deaminase ,chemistry.chemical_compound ,Non-competitive inhibition ,Adenosine Triphosphate ,Adenine nucleotide ,Animal Cells ,Red Blood Cells ,Medicine and Health Sciences ,lcsh:Science ,5'-Nucleotidase ,Mice, Knockout ,Multidisciplinary ,Nucleotides ,Adenine Nucleotides ,Chromatographic Techniques ,AMP deaminase ,Nucleosides ,Animal Models ,Dipyridamole ,Glycosylamines ,Body Fluids ,Adenosine Diphosphate ,Chemistry ,Blood ,Experimental Organism Systems ,Physical Sciences ,Cellular Types ,Anatomy ,Research Article ,Chemical Elements ,Adenosine monophosphate ,Mouse Models ,Research and Analysis Methods ,Equilibrative Nucleoside Transporter 1 ,03 medical and health sciences ,Model Organisms ,Animals ,Cholestenones ,Blood Cells ,Dose-Response Relationship, Drug ,Adenine ,Erythrocyte Membrane ,lcsh:R ,Oxygen transport ,Biology and Life Sciences ,Cell Biology ,Molecular biology ,Adenosine Monophosphate ,Oxygen ,Adenosine diphosphate ,Planar Chromatography ,030104 developmental biology ,chemistry ,Calcium ,lcsh:Q ,Adenosine triphosphate - Abstract
The observation that induced torpor in non-hibernating mammals could result from an increased AMP concentration in circulation led our investigation to reveal that the added AMP altered oxygen transport of erythrocytes. To further study the effect of AMP in regulation of erythrocyte function and systemic metabolism, we generated mouse models deficient in key erythrocyte enzymes in AMP metabolism. We have previously reported altered erythrocyte adenine nucleotide levels corresponding to altered oxygen saturation in mice deficient in both CD73 and AMPD3. Here we further investigate how these Ampd3-/-/Cd73-/- mice respond to the administered dose of AMP in comparison with the control models of single enzyme deficiency and wild type. We found that Ampd3-/-/Cd73-/- mice are more sensitive to AMP-induced hypometabolism than mice with a single enzyme deficiency, which are more sensitive than wild type. A dose-dependent rightward shift of erythrocyte p50 values in response to increasing amounts of extracellular AMP was observed. We provide further evidence for the direct uptake of AMP by erythrocytes that is insensitive to dipyridamole, a blocker for ENT1. The uptake of AMP by the erythrocytes remained linear at the highest concentration tested, 10mM. We also observed competitive inhibition of AMP uptake by ATP and ADP but not by the other nucleotides and metabolites tested. Importantly, our studies suggest that AMP uptake is associated with an erythrocyte ATP release that is partially sensitive to inhibition by TRO19622 and Ca++ ion. Taken together, our study suggests a novel mechanism by which erythrocytes recycle and maintain their adenine nucleotide pool through AMP uptake and ATP release.
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- 2017
34. Metformin In Vitro and In Vivo Increases Adenosine Signaling in Rabbit Corpora Cavernosa
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Paolo Comeglio, Sandra Filippi, Edoardo Mannucci, Giulia Rastrelli, Linda Vignozzi, Ilaria Cellai, Mario Maggi, Annamaria Morelli, and Elena Maneschi
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Male ,medicine.medical_specialty ,Adenosine ,Nitric Oxide Synthase Type III ,Urology ,Endocrinology, Diabetes and Metabolism ,Pharmacology ,Diet, High-Fat ,AMP Deaminase ,Nitric oxide ,Contractility ,chemistry.chemical_compound ,Endocrinology ,Erectile Dysfunction ,In vivo ,Internal medicine ,medicine ,Animals ,Hypoglycemic Agents ,Testosterone ,Enzyme Inhibitors ,business.industry ,Penile Erection ,digestive, oral, and skin physiology ,Receptors, Purinergic P1 ,nutritional and metabolic diseases ,AMP deaminase ,Metformin ,Psychiatry and Mental health ,NG-Nitroarginine Methyl Ester ,Reproductive Medicine ,Mechanism of action ,chemistry ,Rabbits ,medicine.symptom ,business ,Ex vivo ,Penis ,Signal Transduction ,medicine.drug - Abstract
Introduction In subjects with erectile dysfunction responding poorly to sildenafil, metformin was reported to improve erections. Aims The aim of this study is to investigate metformin's mechanism of action on erectile function, particularly focusing on adenosine (ADO) and nitric oxide (NO) signaling in an animal model of high‐fat diet (HFD)‐induced metabolic syndrome. Methods In vitro contractility studies of penile strips. Penile expression of genes related to ADO or NO signaling was also evaluated. Main Outcome Measure In vitro contractility studies were used to investigate the effect of in vivo and ex vivo metformin administration on ADO‐ or acetylcholine (Ach)‐induced relaxation of penile strips from HFD as compared with animals fed a regular diet (RD). Results Expression of ADO receptor type 3 (A 3 R), ADO deaminase (ADA), AMP deaminase type 1 (AMPD1), and 2 (AMPD2) was decreased in HFD as compared with RD. Accordingly, in HFD the ADO relaxant effect was potentiated as compared with RD ( P P P 50 )/IC 50 ratio in RD increased fourfold vs. HFD (RD IC 50 ratio = 13.75 ± 2.96; HFD IC 50 ratio = 2.85 ± 0.52). In corpora cavernosa (CC) from HFD, in vivo metformin (i) normalized A 3 R, ADA, and AMPD1; (ii) further decreased AMPD2; (iii) increased dimethylarginine dimethylamino‐hydrolase; and (iv) partially restored impaired Ach‐induced relaxation. Ex vivo metformin time and dose dependently increased the relaxant effect of ADO in RD. The potentiating effect of metformin on ADO‐induced relaxation was significantly reduced by preincubation with NO synthase inhibitor N ω ‐Nitro‐L‐arginine methyl ester hydrochloride (L‐NAME). Interestingly, in vivo testosterone supplementation in HFD rabbits (i) increased penile expression of endothelial NO synthase and AMPD2 and (ii) restored metformin's potentiating effect on ADO‐induced relaxation up to RD level. Conclusion Metformin in vivo and ex vivo increases ADO signaling in CC, most probably interfering with NO formation and ADO breakdown. Vignozzi L, Filippi S, Comeglio P, Cellai I, Morelli A, Rastrelli G, Maneschi E, Mannucci E, and Maggi M. Metformin in vitro and in vivo increases adenosine signaling in rabbit corpora cavernosa. J Sex Med 2014;11:1694–1708.
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- 2014
35. The Role of Histidine-Proline-Rich Glycoprotein as Zinc Chaperone for Skeletal Muscle AMP Deaminase
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Maria Ranieri-Raggi, Antonio Raggi, and Arthur J. G. Moir
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lcsh:QR1-502 ,chemistry.chemical_element ,Review ,Zinc ,Biology ,Biochemistry ,lcsh:Microbiology ,Metallochaperones ,zinc binding site ,zinc chaperone ,medicine ,Animals ,Humans ,histidine-proline-rich glycoprotein ,Muscle, Skeletal ,AMP deaminase ,Molecular Biology ,Histidine ,Proteins ,Skeletal muscle ,Transport protein ,Protein Transport ,medicine.anatomical_structure ,chemistry ,Chaperone (protein) ,biology.protein ,Molecular Chaperones ,Cysteine - Abstract
Metallochaperones function as intracellular shuttles for metal ions. At present, no evidence for the existence of any eukaryotic zinc-chaperone has been provided although metallochaperones could be critical for the physiological functions of Zn2+ metalloenzymes. We propose that the complex formed in skeletal muscle by the Zn2+ metalloenzyme AMP deaminase (AMPD) and the metal binding protein histidine-proline-rich glycoprotein (HPRG) acts in this manner. HPRG is a major plasma protein. Recent investigations have reported that skeletal muscle cells do not synthesize HPRG but instead actively internalize plasma HPRG. X-ray absorption spectroscopy (XAS) performed on fresh preparations of rabbit skeletal muscle AMPD provided evidence for a dinuclear zinc site in the enzyme compatible with a (μ-aqua)(μ-carboxylato)dizinc(II) core with two histidine residues at each metal site. XAS on HPRG isolated from the AMPD complex showed that zinc is bound to the protein in a dinuclear cluster where each Zn2+ ion is coordinated by three histidine and one heavier ligand, likely sulfur from cysteine. We describe the existence in mammalian HPRG of a specific zinc binding site distinct from the His-Pro-rich region. The participation of HPRG in the assembly and maintenance of skeletal muscle AMPD by acting as a zinc chaperone is also demonstrated.
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- 2014
36. Effect of AMP-Deaminase 3 Knock-Out in Mice on Enzyme Activity in Heart and Other Organs
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Magdalena A. Zabielska, Ryszard T. Smolenski, Krystian Kaletha, Pawel Romaszko, Paul J.R. Barton, Jacek Turyn, Ewa M. Slominska, and Iwona Rybakowska
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medicine.medical_specialty ,Purine nucleoside phosphorylase ,Cre recombinase ,Biochemistry ,AMP Deaminase ,Gene Knockout Techniques ,Mice ,Adenosine deaminase ,Internal medicine ,Genetics ,medicine ,Animals ,chemistry.chemical_classification ,Cardioprotection ,Kidney ,biology ,Chemistry ,Myocardium ,Skeletal muscle ,AMP deaminase ,General Medicine ,Molecular biology ,medicine.anatomical_structure ,Enzyme ,Endocrinology ,Organ Specificity ,biology.protein ,Molecular Medicine ,Gene Deletion - Abstract
Recent findings suggest that inhibition of AMP-deaminase (AMPD) could be effective therapeutic strategy in heart disease associated with cardiac ischemia. To establish experimental model to study protective mechanisms of AMPD inhibition we developed conditional, cardiac specific knock-outs in Cre recombinase system. AMPD3 floxed mice were crossed with Mer-Cre-Mer mice. Tamoxifen was injected to induce Cre recombinase. After two weeks, hearts, skeletal muscle, liver, kidney, and blood were collected and activities of AMPD and related enzymes were analyzed using HPLC-based procedure. We demonstrate loss of more than 90% of cardiac AMPD activity in the heart of AMPD3 -/- mice while other enzymes of nucleotide metabolism such as adenosine deaminase, purine nucleoside phosphorylase were not affected. Surprisingly, activity of AMPD was also reduced in the erythrocytes and in the kidney by 20%–30%. No change of AMPD activity was observed in the skeletal muscle and the liver.
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- 2014
37. Activity of AMP-Regulated Protein Kinase and AMP-Deaminase in the Heart of Mice Fed High-Fat Diet
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Pawel Romaszko, Paulina Żukowska, Marcin Lipiński, Ryszard T. Smolenski, Ewa M. Slominska, and Iwona Rybakowska
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Blood Glucose ,Male ,medicine.medical_specialty ,Energy metabolism ,Peptide ,Diet, High-Fat ,Biochemistry ,AMP Deaminase ,Mice ,Internal medicine ,Genetics ,medicine ,Animals ,Protein kinase A ,chemistry.chemical_classification ,Chemistry ,Activator (genetics) ,Myocardium ,Adenylate Kinase ,Body Weight ,AMPK ,Heart ,AMP deaminase ,High fat diet ,General Medicine ,Endocrinology ,AMPK activity ,Molecular Medicine - Abstract
AMP-regulated protein kinase (AMPK) is involved in numerous regulatory processes and its role in control of cardiac energy metabolism is particularly important. This activity could be affected by AMP-deaminase (AMPD) since substrate of AMPD is AMPK activator. Hearts of male mouse, fed for six weeks with normal or high-fat diet, were fractionated to enrich AMPK activity. Purified fraction was incubated with AMARA peptide for up to 5 minutes and then conversion of AMARA to pAMARA was determined by liquid chromatography-mass spectrometry (LC/MS) using mass detector. Activity of AMPK in heart was 0.038±0.012 pmol/min/mg protein for mice fed high-fat diet and that was not different to control (0.032±0.01 pmol/min/mg protein). We observed change in AMPD activity. It was 5.39±1.5 nmol/mg tissue/min in heart of mice fed high-fat diet while in heart of mice fed low-fat diet it was 2.29±0.32 nmol/mg tissue/min. Data we present indicate that while total AMPK activity is not changed decrease in AMPD activity may affect AMPK signaling in diabetic heart.
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- 2014
38. Straight core structure of DNA replication origins in the mammalian AMPD2 locus
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J. Tavares, Maria Aparecida Fernandez, L F Rodrigues, Q.A. Lima Neto, Fabrícia Gimenes, Adriana Fiorini, Douglas Vinícius Bassalobre de Freitas, Fábio Rogério Rosado, and Fabiana dos Santos Rando
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Genetics ,Base Sequence ,Molecular Sequence Data ,Ter protein ,DNA replication ,Replication Origin ,DNA ,General Medicine ,Computational biology ,Biology ,Pre-replication complex ,Origin of replication ,Biochemistry ,DNA replication origin ,AMP Deaminase ,SeqA protein domain ,Genetic Loci ,Animals ,Origin recognition complex ,DNA supercoil ,Promoter Regions, Genetic - Abstract
Identification of the nucleotide consensus sequence in mammalian replication origins is a difficult and controversial problem. The hypothesis that local DNA topology could be involved in recognition by replication proteins is an exciting possibility. Secondary DNA structures, including intrinsically bent DNA, can be easily detected, and they may indicate a specific pattern in or near mammalian replication origins. This work presents the entire mapping of the intrinsically bent DNA sites (IBDSs), using in silico analysis and a circular permutation assay, of the DNA replication origins oriGNAI3, oriC, oriB, and oriA in the mammalian amplified AMPD2 gene domain. The results show that each origin presents an IBDS that flanks the straight core of these DNA replication sites. In addition, the in silico prediction of the nucleosome positioning reveals a strong indication that the center of an IBDS is localized in a nucleosome-free region (NFR). The structure of each of these curved sites is presented together with their helical parameters and topology. Together, the data that we present here indicate that the oriGNAI3 origin where preferential firing to the replication initiation events in the amplified AMPD2 domain occurs is the only origin that presents a straight, narrow region that is flanked on both sides by two intrinsically bent DNA sites within a short distance (~300 bp); however, all of the origins present at least one IBDS, which is localized in the NFR region. These results indicate that structural features could be implicated in the mammalian DNA replication origin and support the possibility of detecting and characterizing these segments.
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- 2014
39. [Features of influence adenosine, AMP and hyperadrenalinemiya on the immune status, metabolic enzymes of purine nucleotides and the antioxidant defense system]
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S O Tapbergenov, B S Sovetov, and A T Tapbergenov
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0301 basic medicine ,Purine ,Leukocyte migration ,medicine.medical_specialty ,Antioxidant ,Adenosine ,Adrenocortical Hyperfunction ,Adenosine Deaminase ,medicine.medical_treatment ,Glutathione reductase ,General Biochemistry, Genetics and Molecular Biology ,Antioxidants ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Adenosine deaminase ,Internal medicine ,medicine ,Animals ,chemistry.chemical_classification ,biology ,Chemistry ,Glutathione peroxidase ,AMP deaminase ,General Medicine ,Adenosine Monophosphate ,Rats ,030104 developmental biology ,Endocrinology ,030228 respiratory system ,Biochemistry ,biology.protein ,Oxidoreductases ,medicine.drug - Abstract
Administration of a large dose of adrenaline (4 mg/kg 60 min before analysis) increased blood levels of total leukocytes, lymphocytes, decreased T-cell suppressors, leukocyte migration inhibition reaction (LMIR) and NBT test, but increased the level of conjugated dienes (CD). Administration of AMPand adenosine increased levels of total leukocytes, lymphocytes, T- lymphocytes, T-helpers, decreased the level of malondialdehyde (MDA), LMIR, and T-cell suppressors. Sympathetic hyperactivation induced by administration of a large dose of adrenaline (4 mg/kg 60 min before analysis) was accompanied by an increase in heart and liver activities of glutathione peroxidase (GPx), catalase, AMP deaminase (AMPD), and adenosine deaminase (AD). Administration of AMP or adenosine caused a decrease in activities of glutathione reductase (GR), GPx, catalase, a decrease in the MDA level and an increase in activities of AMPD and AD in the heart. In the liver AMP and adenosine also caused a decrease in activities of glutathione reductase (GR), GPx, a decrease in the MDA level and an increase in activities of AMPD and AD. The data obtained suggest that administration of adrenaline, AMP, and adenosine influences activity of enzymes involved in purine nucleotide metabolism. However, in contrast to adrenaline, administration of AMP or adenosine does not provoke stress reaction.Izucheno sostoianie immunnogo statusa, aktivnost' glutationperoksidazy (GPO), glutationreduktazy (GR), katalazy, adenozindezaminazy (AD), AMR-dezaminazy (AMPD), 5'-nukleotidazy (5'N), uroven' malonovogo dial'degida (MDA) i dienovykh kon'iugatov (DK) pri vvedenii zhivotnym adrenalina AMP i adenozina. Adrenalin vyzyval uvelichenie obshchego chisla leĭkotsitov, limfotsitov, snizhenie chisla T-supressorov, RTML i NST, uvelichenie urovnia DK. AMR i adenozin uvelichivali obshchee chislo leĭkotsitov, limfotsitov, T-limfotsitov, T-khelperov, snizhali uroven' MDA, RTML i kolichestvo T-supressorov. Uvelichivaia sootnosheniia aktivnosti AD/AMPD, adrenalin, AMR i adenozin usilivaiut funktsional'nuiu vzaimosviaz' T- i V-zven'ev immuniteta. Vvedenie adrenalina privodilo k uvelicheniiu aktivnosti GPO, katalazy, AD i AMPD v serdtse i pecheni, a AMR i adenozina - k snizheniiu aktivnosti GR, GPO, katalazy, urovnia MDA i povysheniiu aktivnosti AD i AMPD v serdtse. Poluchennye rezul'taty pokazyvaiut, chto pri vvedenii adrenalina, AMR i adenozina proiskhodit uvelichenie aktivnosti fermentov metabolizma purinovykh nukleotidov (AD, AMP) i usilenie funktsional'noĭ vzaimosviazi T- i V-zven'ev immuniteta. V otlichie ot adrenalina, vvedenie AMR i adenozina zhivotnym ne vyzyvaet stressornoĭ reaktsii organizma.
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- 2016
40. Changes in cardiac nucleotide metabolism in Huntington's disease
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Paulina Zukowska, Michal Mielcarek, Marta Toczek, Ryszard T. Smolenski, Mark Isalan, Barbara Kutryb-Zajac, Ewa M. Slominska, and Commission of the European Communities
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0301 basic medicine ,Adenosine ,Adenosine Deaminase ,Cardiomyopathy ,Biochemistry ,AMP Deaminase ,03 medical and health sciences ,Mice ,Adenosine deaminase ,Huntington's disease ,Adenine nucleotide ,Genetics ,medicine ,Animals ,Humans ,Nucleotide ,Pyrophosphatases ,chemistry.chemical_classification ,biology ,adenine nucleotides catabolism ,Myocardium ,Organic Chemistry ,0601 Biochemistry And Cell Biology ,AMP deaminase ,General Medicine ,Metabolism ,medicine.disease ,Molecular biology ,Inosine ,Disease Models, Animal ,030104 developmental biology ,Enzyme ,Huntington Disease ,chemistry ,HD-related cardiomyopathy ,Purine-Nucleoside Phosphorylase ,biology.protein ,Molecular Medicine - Abstract
Huntington's disease (HD) is a monogenic neurodegenerative disorder with a significant peripheral component to the disease pathology. This includes an HD-related cardiomyopathy, with an unknown pathological mechanism. In this study, we aimed to define changes in the metabolism of cardiac nucleotides using the well-established R6/2 mouse model. In particular, we focused on measuring the activity of enzymes that control ATP and other adenine nucleotides in the cardiac pool, including eNTPD, AMPD, e5'NT, ADA, and PNP. We employed HPLC to assay the activities of these enzymes by measuring the concentrations of adenine nucleotide catabolites in the hearts of symptomatic R6/2 mice. We found a reduced activity of AMPD (12.9 ± 1.9 nmol/min/mg protein in control; 7.5 ± 0.5 nmol/min/mg protein in R6/2) and e5'NT (11.9 ± 1.7 nmol/min/mg protein in control; 6.7 ± 0.7 nmol/min/mg protein in R6/2). Moreover, we detected an increased activity of ADA (1.3 ± 0.2 nmol/min/mg protein in control; 5.2 ± 0.5 nmol/min/mg protein in R6/2), while no changes in eNTPD and PNP activities were observed. Analysis of cardiac adenine nucleotide catabolite levels revealed an increased inosine level (0.7 ± 0.01 nmol/mg dry tissue in control; 2.7 ±0.8 nmol/mg dry tissue in R6/2) and a reduced concentration of cardiac adenosine (0.9 ± 0.2 nmol/mg dry tissue in control; 0.2 ± 0.08 nmol/mg dry tissue in R6/2). This study highlights a decreased rate of degradation of cardiac nucleotides in HD mouse model hearts, and an increased capacity for adenosine deamination, that may alter adenosine signaling.
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- 2016
41. Insertion of a knockout-first cassette in Ampd1 gene leads to neonatal death by disruption of neighboring genes expression
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Ying Li, Beisha Tang, Hexiang Peng, Zhengmao Hu, Qiong Liu, Hui Guo, Yongcheng Pan, Jingping Zhao, Kun Xia, Yu Peng, Lusi Zhang, and Jia-Da Li
- Subjects
0301 basic medicine ,Adenosine monophosphate ,Inosine monophosphate ,Mutant ,Biology ,Article ,AMP Deaminase ,Gene Knockout Techniques ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Inosine Monophosphate ,Recombinase ,Animals ,Muscle, Skeletal ,Mice, Knockout ,Regulation of gene expression ,Multidisciplinary ,AMP deaminase ,Phenotype ,Molecular biology ,Adenosine Monophosphate ,Mutagenesis, Insertional ,030104 developmental biology ,Animals, Newborn ,Gene Expression Regulation ,chemistry ,030217 neurology & neurosurgery - Abstract
AMPD1 is an adenosine monophosphate deaminase that catalyzes the deamination of AMP to IMP. To understand the physiological function of AMPD1, we obtained a strain of Ampd1 mutant mice from KOMP repository, which was generated by a knockout-first strategy. An elevated AMP level and almost complete lack of IMP was detected in the skeletal muscle of E18.5 Ampd1tm1a/tm1a mice. However, Ampd1tm1a/tm1a mice died in 2 days postnatally, which was contradicting to previous reports. After removal of the knockout-first cassette and critical exon, mice homozygous for the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d alleles survived to adulthood. RNA-seq analysis indicated that the expression of two neighboring genes, Man1a2 and Nras, were disrupted in the Ampd1tm1a/tm1a mice, but normal in the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d mice. The neonatal lethality phenotype in the Ampd1tm1a/tm1a mice was consistent with the Man1a2-deficient mice. Our results indicated the knockout-first cassette may cause off-target effect by influence the expression of neighboring genes. This study, together with other reports, strongly suggests that removal of targeting cassette by site-specific recombinases is very important for the accurate phenotypic interpretation on mice generated by target mutations.
- Published
- 2016
- Full Text
- View/download PDF
42. Metformin Increases Mitochondrial Energy Formation in L6 Muscle Cell Cultures
- Author
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Raymond S. Ochs and Veeravenkata S. Vytla
- Subjects
Adenosine monophosphate ,Azides ,medicine.medical_specialty ,Time Factors ,Phosphocreatine ,endocrine system diseases ,Cell Survival ,Glucose uptake ,Biology ,Biochemistry ,Cell Line ,chemistry.chemical_compound ,Adenosine Triphosphate ,Internal medicine ,medicine ,Animals ,Hypoglycemic Agents ,Viability assay ,Molecular Biology ,Muscle Cells ,Dose-Response Relationship, Drug ,Uncoupling Agents ,digestive, oral, and skin physiology ,nutritional and metabolic diseases ,AMP deaminase ,Cell Biology ,Adenosine Monophosphate ,Metformin ,Mitochondria, Muscle ,Rats ,Adenosine Diphosphate ,Adenosine diphosphate ,Metabolism ,Endocrinology ,chemistry ,Trypan blue ,Dinitrophenols ,medicine.drug - Abstract
A popular hypothesis for the action of metformin, the widely used anti-diabetes drug, is the inhibition of mitochondrial respiration, specifically at complex I. This is consistent with metformin stimulation of glucose uptake by muscle and inhibition of gluconeogenesis by liver. Yet, mitochondrial inhibition is inconsistent with metformin stimulation of fatty acid oxidation in both tissues. In this study, we measured mitochondrial energy production in intact cells adapting an in vivo technique of phosphocreatine (PCr) formation following energy interruption ("PCr recovery") to cell cultures. Metformin increased PCr recovery from either dinitrophenol (DNP) or azide in L6 cells. We found that metformin alone had no effect on cell viability as measured by total ATP concentration, trypan blue exclusion, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction. However, treatments with low concentrations of DNP or azide reversibly decreased ATP concentration. Metformin increased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction during recovery from either agent. Viability measured by trypan blue exclusion indicated that cells were intact under these conditions. We also found that metformin increased free AMP and, to a smaller extent, free ADP concentrations in cells, an action that was duplicated by a structurally unrelated AMP deaminase inhibitor. We conclude that, in intact cells, metformin can lead to a stimulation of energy formation, rather than an inhibition.
- Published
- 2013
43. Genes for Elite Power and Sprint Performance: ACTN3 Leads the Way
- Author
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David Bishop, Nir Eynon, Peter J. Houweling, Alejandro Lucia, Kathryn N. North, Erik D. Hanson, and Fleur C. Garton
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medicine.medical_specialty ,Genotype ,Nitric Oxide Synthase Type III ,Angiotensinogen ,Physical Therapy, Sports Therapy and Rehabilitation ,Single-nucleotide polymorphism ,Genética humana ,Athletic Performance ,Peptidyl-Dipeptidase A ,Biology ,AMP Deaminase ,Running ,Atleta ,medicine ,Animals ,Humans ,Actinin ,PPAR alpha ,Orthopedics and Sports Medicine ,Muscle Strength ,Muscle, Skeletal ,Gene ,Genetic association ,Genetics ,Polymorphism, Genetic ,Interleukin-6 ,Deporte ,Phenotype ,Sprint ,Elite ,Physical therapy ,Human genome - Abstract
The ability of skeletal muscles to produce force at a high velocity, which is crucial for success in power and sprint performance, is strongly influenced by genetics and without the appropriate genetic make-up, an individual reduces his/her chances of becoming an exceptional power or sprinter athlete. Several genetic variants (i.e. polymorphisms) have been associated with elite power and sprint performance in the last few years and the current paradigm is that elite performance is a polygenic trait, with minor contributions of each variant to the unique athletic phenotype. The purpose of this review is to summarize the specific knowledge in the field of genetics and elite power performance, and to provide some future directions for research in this field. Of the polymorphisms associated with elite power and sprint performance, the a-actinin-3 R577X polymorphism provides the most consistent results. ACTN3 is the only gene that shows a genotype and performance association across multiple cohorts of elite power athletes, and this association is strongly supported by mechanistic data from an Actn3 knockout mouse model. The angiotensin-1 converting enzyme insertion/deletion polymorphism (ACE I/D, registered single nucleotide polymorphism rs]4646994), angiotensinogen (AGT Met235Thr rs699), skeletal adenosine monophosphate deaminase (AMPD1) Gln(Q)12Ter(X) also termed C34T, rs17602729], interleukin-6 (IL-6 -174 G/C, rs1800795), endothelial nitric oxide synthase 3 (NOS3 -786 T/C, rs2070744; and Glu298Asp, rs1799983), peroxisome proliferator-activated receptor-a (PPARA Intron 7 G/C, rs4253778), and mitochondrial uncoupling protein 2 (UCP2 Ala55Val, rs660339) polymorphisms have also been associated with elite power performance, but the findings are less consistent. In general, research into the genetics of athletic performance is limited by a small sample size in individual studies and the heterogeneity of study samples, often including athletes from multiple-difference sporting disciplines. In the future, large, homogeneous, strictly defined elite power athlete cohorts need to be established though multinational collaboration, so that meaningful genome-wide association studies can be performed. Such an approach would provide unbiased identification of potential genes that influence elite athletic performance. 5.320 JCR (2013) Q1, 2/81 Sport sciences UEM
- Published
- 2013
44. AMPD3-deficient mice exhibit increased erythrocyte ATP levels but anemia not improved due to PK deficiency
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Hiroko Morisaki, Jidong Cheng, Masaru Okabe, Keiko Toyama, Takayuki Morisaki, and Masahito Ikawa
- Subjects
Male ,Hemolytic anemia ,Anemia, Hemolytic ,medicine.medical_specialty ,Erythrocytes ,Anemia ,Pyruvate Kinase ,Biology ,AMP Deaminase ,Gene Knockout Techniques ,Mice ,Adenosine Triphosphate ,Internal medicine ,Ribose-Phosphate Pyrophosphokinase ,Genetics ,medicine ,Animals ,Energy charge ,Cells, Cultured ,Mice, Knockout ,Strain (chemistry) ,Erythrocyte fragility ,PK DEFICIENCY ,AMP deaminase ,Cell Biology ,medicine.disease ,Adenosine Monophosphate ,Adenosine Diphosphate ,Mice, Inbred C57BL ,Osmotic Fragility ,Endocrinology ,Biochemistry ,Erythrocyte Count ,Female ,Food Deprivation ,Pyruvate kinase - Abstract
AMP deaminase (AMPD) catalyzes AMP to IMP and plays an important role in energy charge and nucleotide metabolism. Human AMPD3 deficiency is a type of erythrocyte-specific enzyme deficiency found in individuals without clinical symptoms, although an increased level of ATP in erythrocytes has been reported. To better understand the physiological and pathological roles of AMPD3 deficiency, we established a line of AMPD3-deficient [A3(-/-)] mice. No AMPD activity and a high level of ATP were observed in erythrocytes of these mice, similar to human RBC-AMPD3 deficiency, while other characteristics were unremarkable. Next, we created AMPD3 and pyruvate kinase (PK) double-deficient [PKA(-/-,-/-)] mice by mating A3(-/-) mice with CBA-Pk-1slc/Pk-1slc mice [PK(-/-)], a spontaneous PK-deficient strain showing hemolytic anemia. In PKA(-/-,-/-) mice, the level of ATP in red blood cells was increased 1.5 times as compared to PK(-/-) mice, although hemolytic anemia in those animals was not improved. In addition, we observed osmotic fragility of erythrocytes in A3(-/-) mice under fasting conditions. In contrast, the ATP level in erythrocytes was elevated in A3(-/-) mice as compared to the control. In conclusion, AMPD3 deficiency increases the level of ATP in erythrocytes, but does not improve anemia due to PK deficiency and leads to erythrocyte dysfunction.
- Published
- 2012
45. Ability of HMGB1 protein to bind to intrinsically bent and non-bent DNA sites in the AMPD2 gene amplicon
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Maria Aparecida Fernandez, Adriana Fiorini, J. R. Pattaro Júnior, Fábio Rogério Rosado, Q.A. Lima Neto, Vanessa Pinatto Gaspar, K.J.R. Passos, and Douglas Vinícius Bassalobre de Freitas
- Subjects
0301 basic medicine ,DNA Replication ,Replication Origin ,HMGB1 ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,Cricetulus ,Genetics ,Animals ,Binding site ,HMGB1 Protein ,Molecular Biology ,Gene ,Binding Sites ,biology ,DNA replication ,General Medicine ,DNA ,Amplicon ,Molecular biology ,Chromatin ,Cell biology ,Thymine ,DNA-Binding Proteins ,030104 developmental biology ,chemistry ,biology.protein ,Nucleic Acid Conformation ,Protein Binding - Abstract
HMGB-like proteins are architectural chromatin factors, and their function is heavily dependent on their ability to interact with DNA (especially non-canonical DNA structures). HMGB1 is involved in many DNA processes, and dysregulation of HMGB protein expression has profound effects on cellular transcription, resulting in severe developmental defects as well as cancer. During DNA replication, elements that form the origin are still not well defined in metazoans. Sites with A (adenine) or T (thymine) repeats cause intrinsic curvatures in the DNA and are described to be involved in the replication machinery by providing binding sites to replication proteins. As a result, the DNA molecule shows intrinsically bent DNA sites, caused by periodic repeats of 2 or more As/Ts (dA/dT) as well as intrinsically non-bent DNA sites (INBDs), due to a succession of curvatures that cancel each other. In the present study, we mapped 11 INBDSs present in the AMPD2 gene that are related to each replication origin (oriGNAI3, oriC, oriB, and oriA). Following characterization of INBDSs, we tested the ability of HMGB1 to bind to the bent (b1, b2, b4a, b4b, b5, b6, b7, and b8) and non-bent DNA fragments (nb7, nb11, nb1, nb2, nb4, and nb5) via electrophoretic mobility shift assays. All fragments showed efficient binding to HMGB1. However, the non-bent DNA fragments nb2, nb4, and nb5 showed slightly reduced binding efficiency.
- Published
- 2016
46. CL316243 induces phosphatidylinositol 3,4,5-triphosphate production in rat adipocytes in an adenosine deaminase-, pertussis toxin-, or wortmannin-sensitive manner
- Author
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Y Ohsaka and Y Nomura
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0301 basic medicine ,Agonist ,Physiology ,medicine.drug_class ,Adenosine Deaminase ,Phosphatidylinositol Phosphates ,Adrenergic beta-3 Receptor Agonists ,Dioxoles ,Pertussis toxin ,Wortmannin ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Adenosine deaminase ,medicine ,Adipocytes ,Animals ,Phosphatidylinositol ,chemistry.chemical_classification ,biology ,AMP deaminase ,General Medicine ,Molecular biology ,Rats ,Androstadienes ,030104 developmental biology ,Enzyme ,chemistry ,Pertussis Toxin ,030220 oncology & carcinogenesis ,biology.protein - Abstract
The effect of beta(3)-adrenoceptor (beta(3)-AR) agonists on adipocytes treated or not treated with signaling modulators has not been sufficiently elucidated. Using rat epididymal adipocytes (adipocytes) labeled with [(32)P]orthophosphate, we found that treatment with the selective beta(3)-AR agonist CL316243 (CL; 1 microM) induces phosphatidylinositol (PI) 3,4,5-triphosphate (PI[3,4,5]P(3)) production and that this response is inhibited by adenosine deaminase (ADA, an adenosine-degrading enzyme; 2 U/ml), pertussis toxin (PTX, an inactivator of inhibitory guanine-nucleotide-binding protein; 1 microg/ml), or wortmannin (WT, a PI-kinase inhibitor; 3 microM). The results showed that CL induced PI(3,4,5)P(3) production in intact adipocytes and that this production was affected by signaling modulators. Taken together, our findings indicate that CL produces PI(3,4,5)P(3) in an ADA-sensitive, PTX-sensitive, or WT-sensitive manner and will advance understanding of the effect of beta(3)-AR agonists on adipocytes.
- Published
- 2016
47. Host and parasite genetics shape a link between Trypanosoma cruzi infection dynamics and chronic cardiomyopathy
- Author
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Lewis, Michael D., Francisco, Amanda Fortes, Taylor, Martin C., Jayawardhana, Shiromani, and Kelly, John M.
- Subjects
Male ,Mice, Inbred BALB C ,Mice, Inbred C3H ,Myocardium ,Trypanosoma cruzi ,Genes, Protozoan ,Genetic Variation ,Original Articles ,Mice, SCID ,AMP Deaminase ,Host-Parasite Interactions ,Gastrointestinal Tract ,Mice, Inbred C57BL ,Animals ,Original Article ,Chagas Disease ,Female ,Cardiomyopathies - Abstract
Summary Host and parasite diversity are suspected to be key factors in Chagas disease pathogenesis. Experimental investigation of underlying mechanisms is hampered by a lack of tools to detect scarce, pleiotropic infection foci. We developed sensitive imaging models to track Trypanosoma cruzi infection dynamics and quantify tissue‐specific parasite loads, with minimal sampling bias. We used this technology to investigate cardiomyopathy caused by highly divergent parasite strains in BALB/c, C3H/HeN and C57BL/6 mice. The gastrointestinal tract was unexpectedly found to be the primary site of chronic infection in all models. Immunosuppression induced expansion of parasite loads in the gut and was followed by widespread dissemination. These data indicate that differential immune control of T. cruzi occurs between tissues and shows that the large intestine and stomach provide permissive niches for active infection. The end‐point frequency of heart‐specific infections ranged from 0% in TcVI‐CLBR‐infected C57BL/6 to 88% in TcI‐JR‐infected C3H/HeN mice. Nevertheless, infection led to fibrotic cardiac pathology in all models. Heart disease severity was associated with the model‐dependent frequency of dissemination outside the gut and inferred cumulative heart‐specific parasite loads. We propose a model of cardiac pathogenesis driven by periodic trafficking of parasites into the heart, occurring at a frequency determined by host and parasite genetics.
- Published
- 2016
48. Effects of different concentrations of metal ions on degradation of adenosine triphosphate in common carp (Cyprinus carpio) fillets stored at 4°C: An in vivo study
- Author
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Dapeng Li, Longteng Zhang, Na Qin, Yongkang Luo, Jian Lv, and Qingzheng Li
- Subjects
Inosine monophosphate ,Carps ,Metal ions in aqueous solution ,01 natural sciences ,Analytical Chemistry ,chemistry.chemical_compound ,Common carp ,0404 agricultural biotechnology ,Adenosine Triphosphate ,Inosine Monophosphate ,Food Preservation ,Animals ,Carp ,Hypoxanthine ,Ions ,biology ,Chemistry ,010401 analytical chemistry ,Acid phosphatase ,AMP deaminase ,04 agricultural and veterinary sciences ,General Medicine ,biology.organism_classification ,040401 food science ,0104 chemical sciences ,Biochemistry ,Seafood ,Metals ,biology.protein ,bacteria ,Adenosine triphosphate ,Food Science ,Nuclear chemistry - Abstract
The impact of different concentrations of Na + , K + , Ca 2+ , Mg 2+ , Fe 2+ , and Zn 2+ on the degradation of adenosine triphosphate (ATP) and the influence of these ions on the activity of adenosine monophosphate deaminase (AMP-deaminase) and acid phosphatase (ACP) in common carp fillets ( in vivo ) during 4 °C storage was examined. The content of ATP, inosine monophosphate (IMP), and hypoxanthine (Hx), and the activity of AMP-deaminase and ACP were determined. Results indicated that the effects of different concentrations of six kinds of metal ions on AMP-deaminase and ACP were not the same. Na + , K + , Fe 2+ , and Zn 2+ enhanced AMP-deaminase activity, which led to the rapid degradation of ATP and to the generation of a large quantity of IMP within a short time. Ca 2+ and Mg 2+ delayed the change in AMP-deaminase and ACP activity in carp and caused a further delay in the degradation of ATP. Fe 2+ and Zn 2+ inhibited ACP activity, which reduced the decomposition of IMP and the formation of Hx.
- Published
- 2016
49. Maternal fructose drives placental uric acid production leading to adverse fetal outcomes
- Author
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Suzanne M. Scheaffer, Kelle H. Moley, Zeenat Asghar, Noor Al-Hammadi, Alysha Thompson, Maggie M.-Y. Chi, Jessica Saben, and Andrew Cusumano
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Xanthine Oxidase ,medicine.drug_class ,Allopurinol ,Placenta ,030209 endocrinology & metabolism ,Placental insufficiency ,Fructose ,Biology ,Article ,AMP Deaminase ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Pregnancy ,Internal medicine ,medicine ,Animals ,Xanthine oxidase ,Xanthine oxidase inhibitor ,Triglycerides ,2. Zero hunger ,Multidisciplinary ,Fetal Growth Retardation ,medicine.disease ,Placental Insufficiency ,3. Good health ,Uric Acid ,Disease Models, Animal ,Oxidative Stress ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Uric acid ,Female ,Metabolic syndrome ,medicine.drug - Abstract
Maternal metabolic diseases increase offspring risk for low birth weight and cardiometabolic diseases in adulthood. Excess fructose consumption may confer metabolic risks for both women and their offspring. However, the direct consequences of fructose intake per se are unknown. We assessed the impact of a maternal high-fructose diet on the fetal-placental unit in mice in the absence of metabolic syndrome and determined the association between maternal serum fructose and placental uric acid levels in humans. In mice, maternal fructose consumption led to placental inefficiency, fetal growth restriction, elevated fetal serum glucose and triglyceride levels. In the placenta, fructose induced de novo uric acid synthesis by activating the activities of the enzymes AMP deaminase and xanthine oxidase. Moreover, the placentas had increased lipids and altered expression of genes that control oxidative stress. Treatment of mothers with the xanthine oxidase inhibitor allopurinol reduced placental uric acid levels, prevented placental inefficiency and improved fetal weights and serum triglycerides. Finally, in 18 women delivering at term, maternal serum fructose levels significantly correlated with placental uric acid levels. These findings suggest that in mice, excess maternal fructose consumption impairs placental function via a xanthine oxidase/uric acid-dependent mechanism and similar effects may occur in humans.
- Published
- 2016
50. AMPD3 is involved in anthrax LeTx-induced macrophage cell death
- Author
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Jiahuai Han, Yanhai Wang, Sangun Lee, and Sung Ouk Kim
- Subjects
Programmed cell death ,Necrosis ,Cell Survival ,Bacterial Toxins ,Blotting, Western ,Molecular Sequence Data ,Exotoxins ,Mitogen-activated protein kinase kinase ,Polymerase Chain Reaction ,Biochemistry ,AMP Deaminase ,Cell Line ,Anthrax ,Mice ,Drug Discovery ,medicine ,Animals ,Protein kinase A ,Cells, Cultured ,Antigens, Bacterial ,Base Sequence ,Cell Death ,biology ,Macrophages ,Cell Biology ,biology.organism_classification ,Molecular biology ,Bacillus anthracis ,Cell biology ,Cell culture ,Apoptosis ,Tumor necrosis factor alpha ,medicine.symptom ,Research Article ,Biotechnology - Abstract
The responses of macrophages to Bacillus anthracis infection are important for the survival of the host, since macrophages are required for the germination of B. anthracis spores in lymph nodes, and macrophage death exacerbates anthrax lethal toxin (LeTx)-induced organ collapse. To elucidate the mechanism of macrophage cell death induced by LeTx, we performed a genetic screen to search for genes associated with LeTx-induced macrophage cell death. RAW264.7 cells, a macrophage-like cell line sensitive to LeTx-induced death, were randomly mutated and LeTx-resistant mutant clones were selected. AMP deaminase 3 (AMPD3), an enzyme that converts AMP to IMP, was identified to be mutated in one of the resistant clones. The requirement of AMPD3 in LeTx-induced cell death of RAW 264.7 cells was confirmed by the restoration of LeTx sensitivity with ectopic reconstitution of AMPD3 expression. AMPD3 deficiency does not affect LeTx entering cells and the cleavage of mitogen-activated protein kinase kinase (MKK) by lethal factor inside cells, but does impair an unknown downstream event that is linked to cell death. Our data provides new information regarding LeTx-induced macrophage death and suggests that there is a key regulatory site downstream of or parallel to MKK cleavage that controls the cell death in LeTx-treated macrophages.
- Published
- 2011
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