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14 results on '"Nucleobindin"'

1. Expression, purification, and characterization of mouse nesfatin-1 in Escherichia coli.

Author

Xiao C, Liu J, Tang Y, Chen J, Wu X, Bi F, and Zhang J

Subjects
Animals, Escherichia coli genetics, Mice, Nucleobindins, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Calcium-Binding Proteins biosynthesis, Calcium-Binding Proteins chemistry, Calcium-Binding Proteins genetics, Calcium-Binding Proteins isolation & purification, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins chemistry, DNA-Binding Proteins genetics, DNA-Binding Proteins isolation & purification, Escherichia coli chemistry, Escherichia coli metabolism, Gene Expression, Nerve Tissue Proteins biosynthesis, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins genetics, Nerve Tissue Proteins isolation & purification
Abstract

Nesfatin-1 is a newly discovered satiety molecule expressed mainly in the hypothalamic nuclei. It suppresses both short-term and long-term appetite. Six synthetic deoxyoligonucleotides overlapped by PCR encoding nesfatin-1 were cloned into a pET28a vector after the hexa-histidine-tagged multiple cloning sites sequence with an enterokinase recognition site incorporated in-between. The recombinant plasmid was transformed into Escherichia coli strain Rosetta to express the fusion protein, which constituted 27% of the total cell proteins. After purified by Ni-sepharose affinity chromatography, the fusion protein was treated with enterokinase to release nesfatin-1. The nesfatin-1 sample was further purified with reverse-phase high performance liquid chromatography (HPLC), and its molecular weight was determined by mass spectrometry. The biological activities of recombinant nesfatin-1 were also assessed using in vivo animal models. The method described here promises to produce about 8 mg biologically active nesfatin-1 with homogeneity over 98% from 1-L shaking flask culture of E. coli, which can be considered as an easy and cost-effective way to synthesize nesfatin-1., (© 2015 International Union of Biochemistry and Molecular Biology, Inc.)

Published
2017
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2. Calnuc Function in Endosomal Sorting of Lysosomal Receptors.

Author

Larkin H, Costantino S, Seaman MN, and Lavoie C

Subjects
Animals, COS Cells, Calcium-Binding Proteins genetics, Chlorocebus aethiops, DNA-Binding Proteins genetics, HeLa Cells, Humans, Nerve Tissue Proteins genetics, Nucleobindins, Protein Transport, trans-Golgi Network metabolism, Adaptor Proteins, Vesicular Transport metabolism, Calcium-Binding Proteins metabolism, DNA-Binding Proteins metabolism, Endosomes metabolism, Nerve Tissue Proteins metabolism, Receptor, IGF Type 2 metabolism
Abstract

Calnuc is a ubiquitous Ca(2+)-binding protein present on the trans-Golgi network (TGN) and endosomes. However, the precise role of Calnuc in these organelles is poorly characterized. We previously highlighted the role of Calnuc in the transport of LRP9, a new member of a low-density lipoprotein (LDL) receptor subfamily that cycles between the TGN and endosomes. The objective of this study was to explore the role of Calnuc in the endocytic sorting of mannose-6-phosphate receptor (MPR) and Sortilin, two well-characterized lysosomal receptors that transit between the TGN and endosomes. Using biochemical and microscopy assays, we showed that Calnuc depletion [by small interfering RNA (siRNA)] causes the misdelivery to and degradation in lysosomes of cationic-independent mannose-6-phosphate receptor (CI-MPR) and Sortilin due to a defect in the endosomal recruitment of retromers, which are key components of the endosome-to-Golgi retrieval machinery. Indeed, we demonstrated that Calnuc depletion impairs the activation and membrane association of Rab7, a small G protein required for the endosomal recruitment of retromers. Overall, our data indicate a novel role for Calnuc in the endosome-to-TGN retrograde transport of lysosomal receptors through the regulation of Rab7 activity and the recruitment of retromers to endosomes., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Published
2016
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3. Nucleobindin-1 encodes a nesfatin-1-like peptide that stimulates insulin secretion.

Author

Ramesh N, Mohan H, and Unniappan S

Subjects
Amino Acid Sequence, Animals, Calcium-Binding Proteins genetics, Cells, Cultured, DNA-Binding Proteins genetics, Immunoenzyme Techniques, Insulin genetics, Insulin Secretion, Insulin-Secreting Cells cytology, Insulin-Secreting Cells drug effects, Insulinoma drug therapy, Insulinoma genetics, Insulinoma pathology, Mice, Molecular Sequence Data, Nerve Tissue Proteins genetics, Nucleobindins, Protein Precursors genetics, RNA, Messenger genetics, Rats, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Calcium-Binding Proteins metabolism, DNA-Binding Proteins metabolism, Gene Expression Regulation drug effects, Insulin metabolism, Insulin-Secreting Cells metabolism, Insulinoma metabolism, Nerve Tissue Proteins metabolism, Peptide Fragments pharmacology, Protein Precursors metabolism
Abstract

Nesfatin-1 (82 amino acid) is an anorexigenic and insulinotropic peptide encoded in a secreted precursor, nucleobindin-2 (NUCB2). Nucleobindin-1 (NUCB1) is a protein with very high sequence similarity to NUCB2. We hypothesized that a nesfatin-1 like peptide (NLP) is encoded in NUCB1, and this peptide is biologically active. In silico analysis found a signal peptide cleavage site at position 25 (Arginine) and 26 (Valine) preceding the NLP region in NUCB1 sequence, and potential proprotein convertase cleavage sites at Lys-Arg (KR), forming a 77 amino acid NLP. RT-PCR studies found NUCB1 mRNA in both pancreas and MIN6 cells. NUCB1-like immunoreactivity was detected in mouse insulinoma (MIN6) cells, and pancreatic islet beta cells of mice. In order to determine the biological activity of NLP, MIN6 cells were incubated with synthetic rat NLP. NLP (10nM and 100nM) upregulated preproinsulin mRNA expression and insulin secretion at 1h post-incubation. In identical experiments using MIN6 cells, a scrambled peptide based on the NLP sequence did not elicit any effects on preproinsulin mRNA expression or insulin secretion. From this result, it is clear that an intact NLP sequence is required for its biological activity. NLP appears as another endogenous insulinotropic peptide encoded in NUCB1., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Published
2015
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4. Nutrient status modulates the expression of nesfatin-1 encoding nucleobindin 2A and 2B mRNAs in zebrafish gut, liver and brain.

Author

Hatef A, Shajan S, and Unniappan S

Subjects
Amino Acid Sequence, Animals, Food Deprivation, Ghrelin genetics, Ghrelin metabolism, Immunoenzyme Techniques, Molecular Sequence Data, Nucleobindins, Phylogeny, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Tissue Distribution, Zebrafish genetics, Brain metabolism, Calcium-Binding Proteins genetics, Calcium-Binding Proteins metabolism, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Food, Gastrointestinal Tract metabolism, Gene Expression Regulation, Liver metabolism, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Zebrafish metabolism
Abstract

Nesfatin-1 is a naturally occurring, 82-amino acid peptide processed from the precursor nucleobindin 2 (NUCB2), a highly conserved protein among vertebrates. In fish, two isoforms of NUCB2 (NUCB2A and NUCB2B) exist, and nesfatin-1 has been identified in goldfish and Ya fish. We recently reported the presence and appetite suppressing effects of nesfatin-1 in goldfish. The main objectives of this study were to characterize NUCB2 in zebrafish, and determine whether NUCB2 mRNAs are affected by food availability. Tissue distribution of NUCB2A and NUCB2B mRNAs, and NUCB2/nesfatin-1-like immunoreactivity (ir) in the gut of zebrafish were also investigated. In zebrafish, nesfatin-1 region (1-82 amino acids) in NUCB2A is 78% identical to NUCB2B. Both NUCB2A and NUCB2 mRNAs were most abundant in the liver, while less expression was found in other tissues including the brain and gut. NUCB2/nesfatin-1-like immunoreactivity was detected in the mucosal layer cells of zebrafish anterior gastrointestinal tract. NUCB2A and NUCB2B mRNA expression were decreased in the brain of zebrafish 3h after feeding, and after a 7-day food deprivation. Both NUCB2A and NUCB2B mRNAs in the gut were also decreased following 7 days of food deprivation, while NUCB2B mRNA was increased in the liver. Our results provide molecular and functional evidences to support potential anorectic and metabolic roles for endogenous nesfatin-1 in zebrafish. To our knowledge, this is the first report on NUCB2B characterization in vertebrates., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Published
2015
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5. Expression of gastrointestinal nesfatin-1 and gastric emptying in ventromedial hypothalamic nucleus- and ventrolateral hypothalamic nucleus-lesioned rats.

Author

Tian ZB, Deng RJ, Sun GR, Wei LZ, Kong XJ, Ding XL, Jing X, Zhang CP, and Ge YL

Subjects
Animals, Appetite Regulation, Behavior, Animal, Calcium-Binding Proteins genetics, DNA-Binding Proteins genetics, Duodenum innervation, Duodenum metabolism, Eating, Energy Metabolism, Homeostasis, Hyperphagia metabolism, Hyperphagia physiopathology, Hyperphagia psychology, Hypothalamic Area, Lateral surgery, Male, Nerve Tissue Proteins genetics, Nucleobindins, RNA, Messenger metabolism, Rats, Wistar, Time Factors, Ventromedial Hypothalamic Nucleus surgery, Weight Gain, Calcium-Binding Proteins metabolism, DNA-Binding Proteins metabolism, Gastric Emptying, Gastric Mucosa metabolism, Hypothalamic Area, Lateral physiopathology, Nerve Tissue Proteins metabolism, Stomach innervation, Ventromedial Hypothalamic Nucleus physiopathology
Abstract

Aim: To determine the expression levels of gastrointestinal nesfatin-1 in ventromedial hypothalamic nucleus (VMH)-lesioned (obese) and ventrolateral hypothalamic nucleus (VLH)-lesioned (lean) rats that exhibit an imbalance in their energy metabolism and gastric mobility., Methods: Male Wistar rats were randomly divided into a VMH-lesioned group, a VLH-lesioned group, and their respective sham-operated groups. The animals had free access to food and water, and their diets and weights were monitored after surgery. Reverse transcription-polymerase chain reaction and immunostaining were used to analyse the levels of NUCB2 mRNA and nesfatin-1 immunoreactive (IR) cells in the stomach, duodenum, small intestine, and colon, respectively. Gastric emptying was also assessed using a modified phenol red-methylcellulose recovery method., Results: The VMH-lesioned rats fed normal chow exhibited markedly greater food intake and body weight gain, whereas the VLH-lesioned rats exhibited markedly lower food intake and body weight gain. NUCB2/nesfatin-1 IR cells were localised in the lower third and middle portion of the gastric mucosal gland and in the submucous layer of the enteric tract. Compared with their respective controls, gastric emptying was enhanced in the VMH-lesioned rats (85.94% ± 2.27%), whereas the VLH lesions exhibited inhibitory effects on gastric emptying (29.12% ± 1.62%). In the VMH-lesioned rats, the levels of NUCB2 mRNA and nesfatin-1 protein were significantly increased in the stomach and duodenum and reduced in the small intestine. In addition, the levels of NUCB2 mRNA and nesfatin-1 protein in the VLH-lesioned rats were decreased in the stomach, duodenum, and small intestine., Conclusion: Our study demonstrated that nesfatin-1 level in the stomach and duodenum is positively correlated with body mass. Additionally, there is a positive relationship between gastric emptying and body mass. The results of this study indicate that gastrointestinal nesfatin-1 may play a significant role in gastric mobility and energy homeostasis.

Published
2014
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6. Overexpression of CALNUC (Nucleobindin) Increases Agonist and Thapsigargin Releasable Ca2+ Storage in the Golgi

Author

Lin, Ping, Yao, Yong, Hofmeister, Robert, Tsien, Roger Y, and Farquhar, Marilyn Gist

Subjects
Biochemistry and Cell Biology, Medical Physiology, Biomedical and Clinical Sciences, Biological Sciences, Amino Acid Sequence, Animals, Binding Sites, CHO Cells, Calcium, Calcium-Binding Proteins, Calcium-Transporting ATPases, Cloning, Molecular, Cricetinae, DNA-Binding Proteins, Escherichia coli, Golgi Apparatus, Growth Substances, HeLa Cells, Humans, Kinetics, Liver, Molecular Sequence Data, Nerve Tissue Proteins, Nucleobindins, Protein Folding, Protein Structure, Secondary, Rats, Recombinant Fusion Proteins, Recombinant Proteins, Sequence Alignment, Sequence Homology, Amino Acid, Thapsigargin, Transfection, Golgi resident calcium-binding protein, EF-hand, IP3 receptor, SERCA, nucleobindin, Hela Cells, Medical and Health Sciences, Developmental Biology, Biological sciences, Biomedical and clinical sciences
Abstract

We previously demonstrated that CALNUC, a Ca2+-binding protein with two EF-hands, is the major Ca2+-binding protein in the Golgi by 45Ca2+ overlay (Lin, P., H. Le-Niculescu, R. Hofmeister, J.M. McCaffery, M. Jin, H. Henneman, T. McQuistan, L. De Vries, and M. Farquhar. 1998. J. Cell Biol. 141:1515-1527). In this study we investigated CALNUC's properties and the Golgi Ca2+ storage pool in vivo. CALNUC was found to be a highly abundant Golgi protein (3.8 microg CALNUC/mg Golgi protein, 2.5 x 10(5) CALNUC molecules/NRK cell) and to have a single high affinity, low capacity Ca2+-binding site (Kd = 6.6 microM, binding capacity = 1.1 micromol Ca2+/micromol CALNUC). 45Ca2+ storage was increased by 2.5- and 3-fold, respectively, in HeLa cells transiently overexpressing CALNUC-GFP and in EcR-CHO cells stably overexpressing CALNUC. Deletion of the first EF-hand alpha helix from CALNUC completely abolished its Ca2+-binding capability. CALNUC was correctly targeted to the Golgi in transfected cells as it colocalized and cosedimented with the Golgi marker, alpha-mannosidase II (Man II). Approximately 70% of the 45Ca2+ taken up by HeLa and CHO cells overexpressing CALNUC was released by treatment with thapsigargin, a sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) (Ca2+ pump) blocker. Stimulation of transfected cells with the agonist ATP or IP3 alone (permeabilized cells) also resulted in a significant increase in Ca2+ release from Golgi stores. By immunofluorescence, the IP3 receptor type 1 (IP3R-1) was distributed over the endoplasmic reticulum and codistributed with CALNUC in the Golgi. These results provide direct evidence that CALNUC binds Ca2+ in vivo and together with SERCA and IP3R is involved in establishment of the agonist-mobilizable Golgi Ca2+ store.

Published
1999

7. Immunohistochemical Evidence for Glutamatergic Regulation of Nesfatin-1 Neurons in the Rat Hypothalamus

Author

Zehra Minbay, Ozhan Eyigor, Sema Serter Kocoglu, Duygu Gök Yurtseven, Tıp Fakültesi, Bursa Uludağ Üniversitesi/Tıp Fakültesi/Histoloji ve Embriyoloji Anabilim Dalı., Gök, Duygu Yurtseven, Minbay, Zehra, Ezigör, Özhan, ABE-5128-2020, ABC-1475-2020, and AAW-4867-2021

Subjects
Male, Unclassified drug, Immunofluorescence, Expression, nesfatin-1, GABAergic transmission, Hippocampus, Animal tissue, chemistry.chemical_compound, Satiety molecule, Immunoreactivity, 0302 clinical medicine, Food intake, Blood-brain-barrier, rat, hypothalamus, Cell counting, c-Fos, General Neuroscience, Vaccination, Glutamate receptor, Nucleobindin, Pyroglutamyl-Histidyl-Glycine, DNA-Binding Proteins, Immunohistochemistry, 3. Good health, Kainic acid, Hypothalamus, NMDA receptor, Female, medicine.drug, Adult, Agonist, medicine.medical_specialty, Nesfatin 1, medicine.drug_class, Activation, 030209 endocrinology & metabolism, glutamate, AMPA receptor, Neurosciences & neurology, N methyl dextro aspartic acid receptor stimulating agent, Glutamatergic synapse, Article, lcsh:RC321-571, 03 medical and health sciences, Glutamatergic, Protein phosphorylation, Internal medicine, medicine, Animal model, 6 cyano 7 nitro 2,3 quinoxalinedione, Neurotransmitter, Animal experiment, Synaptic transmission, lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry, N methyl dextro aspartic acid receptor, Neurosciences, Kainate, Energy metabolism, Nonhuman, Immunofluorescence test, Dizocilpine, AMPA receptor agonist, Endocrinology, chemistry, nervous system, Receptor subunits, Protein expression, Ultrastructural-localization, Subunit messenger-RNAs, Controlled study, 030217 neurology & neurosurgery
Abstract

Koçoğlu, Sema Serter (Balikesir Author), Nesfatin-1, identified as an anorexigenic peptide, regulates the energy metabolism by suppressing food intake. The majority of nesfatin-1-synthesizing neurons are concentrated in various hypothalamic nuclei, especially in the supraoptic (SON), arcuate (ARC) and paraventricular nuclei (PVN). We tested the hypothesis that the glutamatergic system regulates nesfatin-1 neurons through glutamate receptors. Therefore, the first aim of the proposed studies was to examine effects of different glutamate agonists in the activation of nesfatin-1 neurons using c-Fos double immunohistochemical labeling. Experimental groups were formed containing male and female rats which received intraperitoneal injections of glutamate agonists kainic acid, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) while the control rats received vehicle. The significant increase in the number of c-Fos-expressing nesfatin-1 neurons after agonist injections were observed both in female and male subjects and some of these effects were found to be sexually dimorphic. In addition, treatment with specific glutamate antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or dizocilpine (MK-801) before each of the three agonist injections caused a statistically significant reduction in the number of activated nesfatin-1 neurons in the hypothalamic nuclei including supraoptic, paraventricular and arcuate nuclei. The second aim of the study was to determine the expression of glutamate receptor subunit proteins in the nesfatin-1 neurons by using a double immunofluorescence technique. The results showed that the glutamate receptor subunits, which may form homomeric or heteromeric functional receptor channels, were expressed in the nesfatin-1 neurons. In conclusion, the results of this study suggest that nesfatin-1 neurons respond to glutamatergic signals in the form of neuronal activation and that the glutamate receptors that are synthesized by nesfatin-1 neurons may participate in the glutamatergic regulation of these neurons.

Published
2020
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8. Nucleobindins: Bioactive precursor proteins encoding putative endocrine factors?

Author

Gonzalez, Ronald, Mohan, Haneesha, and Unniappan, Suraj

Subjects
*BIOACTIVE compounds, *DNA-binding proteins, *CALCIUM, *G proteins, *BRAIN stem, *LABORATORY rats
Abstract

Abstract: The nucleobindins, nucleobindin 1 (NUCB1) and nucleobindin 2 (NUCB2), are homologous multidomain calcium and DNA binding proteins. NUCB1 is a well-characterized Golgi protein found within the rat pituitary, liver and kidney with functions related to immunity, calcium homeostasis and G protein signaling. NUCB2 is found both in the hypothalamus and brain stem centers, as well as peripherally in the digestive tract. Renewed interest in the nucleobindins has been sparked by the recent discovery of nesfatin-1, an endocrine factor post-translationally processed from the N-terminal of NUCB2. Nesfatin-1 has quickly established itself as a novel regulator of appetite, insulin secretion, energy homeostasis and reproduction with important consequences to the etiology of metabolic diseases including diabetes and obesity. The discovery of nesfatin-1 and it endocrine functions attracted more attention to the nucleobindins that are already known to have important intracellular functions. From the sequence information available, it is possible that nucelobindins itself or nesfatin-1 like peptides within the NUCB1 could also elicit nesfatin-1-like biological functions. The research on nesfatin-1 in last 5years further adds to the importance of nucleobindins as potential endocrine precursors. This review aims to summarize some of the most recent findings on the functional significance of NUCB1, NUCB2, as well as encoded proteins and highlights the questions that remain unanswered. [Copyright &y& Elsevier]

Published
2012
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9. The role of centrally injected nesfatin-1 on cardiovascular regulation in normotensive and hypotensive rats

Author

Ilker Arican, Leman Gizem Erkan, Ozge Avsar, Gokcen Guvenc, Vahide Savci, Murat Yalcin, Duygu Udum Kucuksen, Mustafa Sertac Yilmaz, Burcin Altinbas, Uludağ Üniversitesi/Tıp Fakültesi/Tıbbi Farmakoloji Anabilim Dalı., Uludağ Üniversitesi/Veterinerlik Fakültesi/Fizyoloji Anabilim Dalı., Uludağ Üniversitesi/Veterinerlik Fakültesi/Biyokimya Anabilim Dalı., Uludağ Üniversitesi/Veterinerlik Fakültesi/Anatomi Anabilim Dalı., Yılmaz, Mustafa Sertaç, Altınbaş, Burçin, Güvenç, Gökçen, Erkan, Leman Gizem, Avşar, Özge, Savcı, Vahide, Küçükşen, Duygu Udum, Arıcan, İlker, Yalçın, Murat, AAH-1571-2021, AAR-6815-2021, AAH-5167-2021, AAG-7518-2021, and AAG-6956-2021

Subjects
Tachycardia, Male, Vasopressin, Unclassified drug, Physiology, Hemorrhagic hypotension, Pressor response, Blood Pressure, Catecholamine blood level, Rats, Sprague-Dawley, Catecholamines, Heart Rate, Paraventricular nucleus, Renin, Vasopressin derivative, Priority journal, Neurons, Sprague Dawley rat, Brain, Nucleobindin, Pyroglutamyl-Histidyl-Glycine, DNA-Binding Proteins, Cardiovascular effect, Mean arterial pressure, Neuroendocrine, Blood, Hemorrhagic shock, Nesfatin-1, Catecholamine, medicine.symptom, Hypotension, medicine.drug, Bradycardia, medicine.medical_specialty, Vasopressin blood level, Nesfatin 1, Vasopressins, DNA binding proteins, Nerve protein, Hemorrhage, Nerve Tissue Proteins, Neurosciences & neurology, Pathophysiology, Article, Cellular and Molecular Neuroscience, Internal medicine, Renin–angiotensin system, Heart rate, medicine, Animals, Nucleobindins, Animal experiment, Drug effects, Animal, Disease model, Endocrine and Autonomic Systems, business.industry, Bleeding, Calcium-Binding Proteins, Neurosciences, Nonhuman, Rats, Neuropeptide, Disease Models, Animal, Endocrinology, Blood pressure, Sprague-dawley, Rat, Neurology (clinical), Calcium binding proteins, business, Controlled study, Complication, Central Nervous System Agents
Abstract

This study investigated the cardiovascular effects of nesfatin-1 in normotensive rats and animals subjected to hypotensive hemorrhage. Hemorrhagic hypotension was induced by withdrawal 2 mL blood/100 g body weight over a period of 10 min. Acute hemorrhage led to a severe and long-lasting decrease in mean arterial pressure (MAP) and heart rate (HR). Intracerebroventricularly (i.c.v.) administered nesfatin-1 (100 pmol) increased MAP in both normotensive and hemorrhaged rats. Nesfatin-1 also caused bradycardia in normotensive and tachycardia in hemorrhaged rats. Centrally injected nesfatin-1 (100 pmol, i.c.v.) also increased plasma catecholamine, vasopressin and renin concentrations in control animals and potentiated the rise in all three cardiovascular mediators produced by hemorrhage. These findings indicate that centrally administered nesfatin-1 causes a pressor response in conscious normotensive and hemorrhaged rats and suggest that enhanced sympathetic activity and elevated vasopressin and renin concentrations mediate the cardiovascular effects of the peptide.

Published
2015

10. The effect of chronic peripheral nesfatin-1 application on blood pressure in normal and chronic restraint stressed rats: Related with circulating level of blood pressure regulators

Author

Ceylan Ayada, Sebahat Turgut, Günfer Turgut, and Zuhal Güçlü

Subjects
immobilization stress, Male, hydrocortisone blood level, Endothelial nitric oxide synthase (eNOS), Physiology, nerve protein, Blood Pressure, angiotensin II, Wistar rat, Cortisol, angiotensin converting enzyme 2, Renin-Angiotensin System, chemistry.chemical_compound, renin angiotensin aldosterone system, Medicine, rat, animal, physiological stress, Aldosterone, Endothelin-1, blood pressure regulation, General Medicine, gene expression regulation, peptide, Peripheral, unclassified drug, DNA-Binding Proteins, medicine.anatomical_structure, Peripheral nervous system, Angiotensin-converting enzyme 2, Nesfatin-1, blood sampling, nesfatin 1, Renin-angiotensin-aldosterone system (RAAS), medicine.medical_specialty, blood pressure meter, hypertension, Nitric Oxide Synthase Type III, animal experiment, Biophysics, hormone blood level, Enzyme-Linked Immunosorbent Assay, Nerve Tissue Proteins, aldosterone blood level, Article, Gene Expression Regulation, Enzymologic, enzyme blood level, body weight, Stress, Physiological, Internal medicine, Nos3 protein, rat, nucleobindin, Nucleobindins, Animals, endothelin 1, controlled study, hydrocortisone, Rats, Wistar, chronic stress, endothelial nitric oxide synthase, aldosterone, nonhuman, business.industry, animal model, rat model, Calcium-Binding Proteins, calcium binding protein, medicine.disease, Angiotensin II, DNA binding protein, enzyme linked immunosorbent assay, Rats, Endocrinology, Blood pressure, angiotensinogen, chemistry, Pathophysiology of hypertension, drug effects, business, metabolism, Homeostasis
Abstract

Nesfatin is a peptide secreted by peripheral tissues, central and peripheral nervous system. It is involved in the regulation of homeostasis. Although the effects of nesfatin-1 on nutrition have been studied widely in the literature, the mechanisms of nesfatin-1 action and also relations with other physiological parameters are still not clarified well. We aimed to investigate the effect of peripheral chronic nesfatin-1 application on blood pressure regulation in normal and in rats exposed to restraint immobilization stress. In our study, three month-old male Wistar rats were used. Rats were divided into 4 groups as Control, Stress, Control+Nesfatin-1, Nesfatin-1+Stress. Angiotensinogen, angiotensin converting enzyme 2, angiotensin II, endothelin-1, endothelial nitric oxide synthase, aldosterone, cortisol, nesfatin-1 levels were determined in plasma samples by ELISA. Our results have shown that chronic peripheral nesfatin-1 administration increases blood pressure in normal and in rats exposed to chronic restraint stress. Effect of nesfatin-1 on circulating level of angiotensinogen, angiotensin converting enzyme 2, angiotensin II, endothelin-1, endothelial nitric oxide synthase, aldosterone and cortisol has been identified. We can conclude that elevated high blood pressure after chronic peripheral nesfatin-1 administration in rats exposed to chronic restraint stress may be related to decreased plasma level of endothelial nitric oxide synthase concentration.

Published
2015

11. The effect of nesfatin-1 on heart L-type Ca2+ channel alpha 1c subunit in rats subjected to chronic restraint stress

Author

Sebahat Turgut, Ceylan Ayada, and Günfer Turgut

Subjects
Male, Cardiac failure, nerve protein, nesfatin-1, Wistar rat, Muscle hypertrophy, L-type Ca2+ channel alpha 1c subunit, Materials Chemistry, animal, rat, Ca channel, geography.geographical_feature_category, medicine.diagnostic_test, exercise, Chemistry, Forestry, Islet, Peripheral, DNA-Binding Proteins, L-type Ca2+ channel α1c subunit, mental stress, Nesfatin-1, Restraint stress, Restraint, Physical, Economics and Econometrics, medicine.medical_specialty, Calcium Channels, L-Type, Protein subunit, Nerve Tissue Proteins, Western blot, heart, Chronic restraint stress, Internal medicine, nucleobindin, Media Technology, medicine, L-type Ca2+ channel ?1c subunit, calcium channel L type, Animals, Rats, Wistar, Beta (finance), geography, Myocardium, Calcium-Binding Proteins, chronic restraint stress, heart muscle, calcium binding protein, DNA binding protein, Rats, Endocrinology, Cacna1c protein, rat, drug effects, cardiac failure, metabolism, Stress, Psychological
Abstract

WOS: 000357840200008, PubMed: 25924643, Background: Elevated L-type Ca2+ channel expression level increases Ca2+ influx. This can cause hypertrophy and pathological remodeling of the heart especially under stress conditions. Nesfatin-1 can activate hypothalamic L, P and Q type Ca2+ channels and increase insulin secretion in pancreatic islet beta cells via activation of L-type Ca2+ channels. On the other hand, the effect of nesfatin-1 on cardiac L-type Ca2+ channels has not been studied yet. Objectives: We aimed to identify the effect of peripheral chronic nesfatin-1 application on cardiac L-type Ca2+ channel ale subunit expression level in normal rats and those subjected to chronic restraint stress. Methods: Three-month aged VVistar albino rats were randomly divided into 4 groups (n = 7) as Control, Stress, Control+Nesfatin-1, and Nesfatin-1+Stress. Rats in groups subjected to restraint stress were placed in a specially built size-manipulable cabin for 2 h/day (between 10:00 and 12:00 a.m.) for 10 consecutive days without allowing water and food intake. Nesfatin-1 segment (0.25 nrnol/g bw intraperitoneally) was applied during the 10 consecutive days. Western blot analyses were performed to determine the expression level of L-type Ca2+ channel alpha 1c subunit protein in rat cardiac extracts. Results: Cardiac L-type Ca2+ channel alpha 1c subunit protein expression levels were increased significantly after chronic peripheral nesfatin-1 application in rats subjected to restraint stress (p = 0.032). Conclusion: We can conclude that nesfatin-1 can cause cardiac failures during clinical treatments by elevating cardiac L-type Ca2+ channel alpha 1c subunit protein expression level (Fig. 2, Ref. 26). Text in PDF www.elis.sk., Pamukkale UniversityPamukkale University [2011SBE005], This study was supported by Pamukkale University Research Fund (Project no: 2011SBE005). We are grateful to Technical Assistant Zuhal Guclu from Pamukkale University, Experimental Animal Research Unit, Denizli, Turkey, for her excellent technical help.

Published
2015

12. Interaction of the protein nucleobindin with Gαi2, as revealed by the yeast two-hybrid system

Author

Naoki Mochizuki, Yoshiyuki Kanai, Masahiko Hibi, and Paul A. Insel

Subjects
Signal peptide, Leucine zipper, DNA, Complementary, G protein, Recombinant Fusion Proteins, Two-hybrid screening, Molecular Sequence Data, Biophysics, Nerve Tissue Proteins, GTP-binding protein, Saccharomyces cerevisiae, Biology, Transfection, Biochemistry, DNA-binding protein, Cell Line, Mice, GTP-binding protein regulators, GTP-Binding Proteins, Structural Biology, Calcium-binding protein, Heterotrimeric G protein, Chlorocebus aethiops, Genetics, Animals, Humans, Nucleobindins, Growth Substances, Molecular Biology, DNA Primers, Gene Library, Glutathione Transferase, B-Lymphocytes, Base Sequence, Calcium-Binding Proteins, Yeast two-hybrid, Antibodies, Monoclonal, Cell Biology, beta-Galactosidase, DNA-Binding Proteins, Nucleobindin, Signal Transduction
Abstract

The heterotrimeric G protein, G alpha i2, transduces signals from seven membrane spanning receptors to effectors such as adenylyl cyclase and ion channels. The purpose of this study was to identify these or other cellular proteins that interact with G alpha i2 by use of the yeast two-hybrid system. A human B cell cDNA library was screened by this system using full length G alpha i2. Four positive colonies were obtained. Two of the four were identified as nucleobindin, a calcium binding protein and a putative antigen to which anti-nuclear antibodies are generated in mice with a disorder that resembles systemic lupus erythematosus. Nucleobindin has a leucine zipper, EF hands, and a signal peptide sequence and is thought to localize to the nucleus as well as being secreted. The specificity of intehraction between G alpha i2 and nucleobindin was confirmed by an in vitro binding assay using recombinant proteins. Transfection of G alpha i2 and nucleobindin in COS cells increased G alpha i2 expression relative to cells transfected with G alpha i2 and mock vector. Our results indicate that the yeast two-hybrid system provides a means to identify novel proteins that interact with G alpha proteins. Nucleobindin appears to represent one of those proteins.

Published
1995
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13. Cellular distribution, regulated expression, and functional role of the anorexigenic peptide, NUCB2/nesfatin-1, in the testis

Author

Francisco Ruiz-Pino, Manuel Tena-Sempere, María Jesús Vázquez, David Garcia-Galiano, Antonio Romero-Ruiz, Francisco Gaytan, Miguel A. Sánchez-Garrido, Juan M. Castellano, Carlos Dieguez, Rafael Pineda, Susana Sangiao-Alvarellos, Leonor Pinilla, T. Ilhan, Uludağ Üniversitesi/Veteriner Fakültesi/Histoloji ve Embriyoloji Anabilim Dalı., İlhan, Tuncay, and AAH-8859-2021

Subjects
Luteinizing hormone, Male, Cellular distribution, Aging, Unclassified drug, Mouse, Messenger rna, Protein function, Adipose tissue, Energy homeostasis, Animal tissue, Testosterone secretion, Mice, 0302 clinical medicine, Endocrinology, Satiety molecule, Leydig cell, Paraventricular nucleus, Maturation, Testis, Testosterone, Sexual Maturation, Insulin-secretion, Endocrinology & metabolism, Priority journal, 0303 health sciences, Leydig Cells, Sertoli cell, Nucleobindin 2, Chorionic gonadotropin, DNA-Binding Proteins, medicine.anatomical_structure, Models, Animal, Nesfatin-1, Animal cell, Human, endocrine system, medicine.medical_specialty, Ex vivo study, Nesfatin 1, Neuropeptide, 030209 endocrinology & metabolism, Nerve Tissue Proteins, Biology, Article, Ribonucleic-acid expression, 03 medical and health sciences, Diet restriction, Internal medicine, Metabolic regulation, Upregulation, Energy-balance, medicine, Nucleobindin, Pyroglutamyl-Histidyl-Glycine, SHU 9119, Animals, Humans, Nucleobindins, Animal experiment, Progenitor cell, Human tissue, Rats, Wistar, 030304 developmental biology, Messenger RNA, Orexin-a, Puberty, Calcium-Binding Proteins, Leydig-cells, Nonhuman, Rats, Mice, Inbred C57BL, Human cell, Gene Expression Regulation, Rat testis, Protein expression, Rat, Energy Metabolism, Controlled study
Abstract

Nesfatin-1, product of the precursor NEFA/nucleobindin2 (NUCB2), was initially identified as anorectic hypothalamic neuropeptide, acting in a leptin-independent manner. In addition to its central role in the control of energy homeostasis, evidence has mounted recently that nesfatin-1 is also produced in peripheral metabolic tissues, such as pancreas, adipose, and gut. Moreover, nesfatin-1 has been shown to participate in the control of body functions gated by whole-body energy homeostasis, including puberty onset. Yet, whether, as is the case for other metabolic neuropeptides, NUCB2/nesfatin-1 participates in the direct control of gonadal function remains unexplored. Wedocument here for the first time the expression ofNUCB2mRNAin rat, mouse, and human testes, where NUCB2/nesfatin-1 protein was identified in interstitial mature Leydig cells. Yet in rats, NUCB2/nesfatin-1becameexpressed in Sertoli cellsuponLeydig cell eliminationandwas also detected in Leydig cell progenitors. Although NUCB2 mRNA levels did not overtly change in rat testis during pubertal maturation and after short-term fasting, NUCB2/nesfatin-1 content significantly increased along the puberty-to-adult transition and was markedly suppressed after fasting. In addition, testicular NUCB2/nesfatin-1 expression was up-regulated by pituitary LH, because hypophysectomy decreased, whereas human choriogonadotropin (super-agonist of LH receptors) replacement enhanced, NUCB2/nesfatin-1 mRNA and peptide levels. Finally, nesfatin-1 increased human choriogonadotropin-stimulated testosterone secretion by rat testicular explants ex vivo. Our data are the first to disclose the presence and functional role of NUCB2/nesfatin-1 in the testis, where its expression is regulated by developmental, metabolic, and hormonal cues as well as by Leydig cell-derived factors. Our observations expand the reproductive dimension of nesfatin-1, which may operate directly at the testicular level to link energy homeostasis, puberty onset, and gonadal function. Spanish Government - BFU 2008-00984 / BFU 2011-25021 / PI042082 European Commission - DEER FP7-ENV-2007-1 Junta de Andalucia European Commission - P08-CVI-03788

Published
2012

14. Overexpression of CALNUC (nucleobindin) increases agonist and thapsigargin releasable Ca2+ storage in the Golgi

Author

Yong Yao, Roger Y. Tsien, Robert Hofmeister, Marilyn G. Farquhar, and Ping Lin

Subjects
Secondary, Protein Folding, Sarcoplasm, Golgi Apparatus, Sequence Homology, EF-hand, Medical and Health Sciences, Protein Structure, Secondary, chemistry.chemical_compound, 0302 clinical medicine, Calcium-binding protein, Cricetinae, Cloning, Molecular, Growth Substances, 0303 health sciences, Biological Sciences, Recombinant Proteins, Cell biology, DNA-Binding Proteins, Amino Acid, Liver, symbols, Thapsigargin, Protein Structure, SERCA, Recombinant Fusion Proteins, Molecular Sequence Data, IP3 receptor, Nerve Tissue Proteins, CHO Cells, Calcium-Transporting ATPases, Biology, Transfection, Vaccine Related, 03 medical and health sciences, symbols.namesake, nucleobindin, Escherichia coli, Animals, Humans, Nucleobindins, Amino Acid Sequence, 030304 developmental biology, Binding Sites, Sequence Homology, Amino Acid, Endoplasmic reticulum, Calcium-Binding Proteins, Molecular, Cell Biology, Golgi apparatus, Inositol trisphosphate receptor, Molecular biology, Rats, Calcium ATPase, Kinetics, chemistry, Hela Cells, Golgi resident calcium-binding protein, Calcium, Generic health relevance, Sequence Alignment, 030217 neurology & neurosurgery, HeLa Cells, Regular Articles, Cloning, Developmental Biology
Abstract

We previously demonstrated that CALNUC, a Ca2+-binding protein with two EF-hands, is the major Ca2+-binding protein in the Golgi by 45Ca2+ overlay (Lin, P., H. Le-Niculescu, R. Hofmeister, J.M. McCaffery, M. Jin, H. Henneman, T. McQuistan, L. De Vries, and M. Farquhar. 1998. J. Cell Biol. 141:1515-1527). In this study we investigated CALNUC's properties and the Golgi Ca2+ storage pool in vivo. CALNUC was found to be a highly abundant Golgi protein (3.8 microg CALNUC/mg Golgi protein, 2.5 x 10(5) CALNUC molecules/NRK cell) and to have a single high affinity, low capacity Ca2+-binding site (Kd = 6.6 microM, binding capacity = 1.1 micromol Ca2+/micromol CALNUC). 45Ca2+ storage was increased by 2.5- and 3-fold, respectively, in HeLa cells transiently overexpressing CALNUC-GFP and in EcR-CHO cells stably overexpressing CALNUC. Deletion of the first EF-hand alpha helix from CALNUC completely abolished its Ca2+-binding capability. CALNUC was correctly targeted to the Golgi in transfected cells as it colocalized and cosedimented with the Golgi marker, alpha-mannosidase II (Man II). Approximately 70% of the 45Ca2+ taken up by HeLa and CHO cells overexpressing CALNUC was released by treatment with thapsigargin, a sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) (Ca2+ pump) blocker. Stimulation of transfected cells with the agonist ATP or IP3 alone (permeabilized cells) also resulted in a significant increase in Ca2+ release from Golgi stores. By immunofluorescence, the IP3 receptor type 1 (IP3R-1) was distributed over the endoplasmic reticulum and codistributed with CALNUC in the Golgi. These results provide direct evidence that CALNUC binds Ca2+ in vivo and together with SERCA and IP3R is involved in establishment of the agonist-mobilizable Golgi Ca2+ store.

Published
1999

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