91 results on '"Yong Juan Zhao"'
Search Results
2. A non-internalised CD38-binding radiolabelled single-domain antibody fragment to monitor and treat multiple myeloma
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Elodie Duray, Margaux Lejeune, Frederic Baron, Yves Beguin, Nick Devoogdt, Ahmet Krasniqi, Yoline Lauwers, Yong Juan Zhao, Matthias D’Huyvetter, Mireille Dumoulin, and Jo Caers
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Multiple myeloma ,CD38 ,Theranostic ,Single-domain antibody ,Nanobody ,Radio-immunotherapy ,Diseases of the blood and blood-forming organs ,RC633-647.5 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Antibody-based therapies targeting CD38 are currently used as single agents as well as in combination regimens for multiple myeloma, a malignant plasma cell disorder. In this study, we aimed to develop anti-CD38 single-domain antibodies (sdAbs) that can be used to trace CD38+ tumour cells and subsequently used for targeted radionuclide therapy. SdAbs are derived from Camelidae heavy-chain antibodies and have emerged as promising theranostic agents due to their favourable pharmacological properties. Methods Four different anti-CD38 sdAbs were produced, and their binding affinities and potential competition with the monoclonal antibody daratumumab were tested using biolayer interferometry. Their binding kinetics and potential cell internalisation were further studied after radiolabelling with the diagnostic radioisotope Indium-111. The resulting radiotracers were evaluated in vivo for their tumour-targeting potential and biodistribution through single-photon emission computed tomography (SPECT/CT) imaging and serial dissections. Finally, therapeutic efficacy of a lead anti-CD38 sdAb, radiolabelled with the therapeutic radioisotope Lutetium-177, was evaluated in a CD38+ MM xenograft model. Results We retained anti-CD38 sdAb #2F8 as lead based on its excellent affinity and superior stability, the absence of competition with daratumumab and the lack of receptor-mediated internalisation. When intravenously administered to tumour-xenografted mice, radiolabelled sdAb #2F8 revealed specific and sustained tumour retention with low accumulation in other tissues, except kidneys, resulting in high tumour-to-normal tissue ratios. In a therapeutic setting, myeloma-bearing mice received three consecutive intravenous administrations of a high (18.5 MBq) or a low radioactive dose (9.3 MBq) of 177Lu-DTPA-2F8 or an equal volume of vehicle solution. A dose-dependent tumour regression was observed, which translated into a prolonged median survival from 43 days for vehicle-treated mice, to 62 days (p = 0.027) in mice receiving the low and 65 days in mice receiving the high (p = 0.0007) radioactive dose regimen, respectively. Conclusions These results highlight the theranostic potential of radiolabelled anti-CD38 sdAbs for the monitoring and treatment of multiple myeloma.
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- 2021
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3. The complex structure of GRL0617 and SARS-CoV-2 PLpro reveals a hot spot for antiviral drug discovery
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Ziyang Fu, Bin Huang, Jinle Tang, Shuyan Liu, Ming Liu, Yuxin Ye, Zhihong Liu, Yuxian Xiong, Wenning Zhu, Dan Cao, Jihui Li, Xiaogang Niu, Huan Zhou, Yong Juan Zhao, Guoliang Zhang, and Hao Huang
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Science - Abstract
The SARS-CoV-2 papain-like protease (PLpro) is of interest as a drug target. Here, the authors identify GRL0617 as a PPI (protein–protein interaction) inhibitor of SARS-CoV-2 PLpro that inhibits its deISGylating activity and present the mechanism of action of the compound through the GRL0617-bound PLpro crystal structure and NMR studies.
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- 2021
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4. Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate
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Wan Hua Li, Ke Huang, Yang Cai, Qian Wen Wang, Wen Jie Zhu, Yun Nan Hou, Sujing Wang, Sheng Cao, Zhi Ying Zhao, Xu Jie Xie, Yang Du, Chi-Sing Lee, Hon Cheung Lee, Hongmin Zhang, and Yong Juan Zhao
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SARM1 ,fluorescent probes ,base-exchange ,allosteric inhibitors ,CryoEM structure ,covalent inhibitors ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
SARM1 regulates axonal degeneration through its NAD-metabolizing activity and is a drug target for neurodegenerative disorders. We designed and synthesized fluorescent conjugates of styryl derivative with pyridine to serve as substrates of SARM1, which exhibited large red shifts after conversion. With the conjugates, SARM1 activation was visualized in live cells following elevation of endogenous NMN or treatment with a cell-permeant NMN-analog. In neurons, imaging documented mouse SARM1 activation preceded vincristine-induced axonal degeneration by hours. Library screening identified a derivative of nisoldipine (NSDP) as a covalent inhibitor of SARM1 that reacted with the cysteines, especially Cys311 in its ARM domain and blocked its NMN-activation, protecting axons from degeneration. The Cryo-EM structure showed that SARM1 was locked into an inactive conformation by the inhibitor, uncovering a potential neuroprotective mechanism of dihydropyridines.
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- 2021
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5. A Cell-Permeant Mimetic of NMN Activates SARM1 to Produce Cyclic ADP-Ribose and Induce Non-apoptotic Cell Death
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Zhi Ying Zhao, Xu Jie Xie, Wan Hua Li, Jun Liu, Zhe Chen, Ben Zhang, Ting Li, Song Lu Li, Jun Gang Lu, Liangren Zhang, Li-he Zhang, Zhengshuang Xu, Hon Cheung Lee, and Yong Juan Zhao
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Science - Abstract
Summary: SARM1, an NAD-utilizing enzyme, regulates axonal degeneration. We show that CZ-48, a cell-permeant mimetic of NMN, activated SARM1 in vitro and in cellulo to cyclize NAD and produce a Ca2+ messenger, cADPR, with similar efficiency as NMN. Knockout of NMN-adenylyltransferase elevated cellular NMN and activated SARM1 to produce cADPR, confirming NMN was its endogenous activator. Determinants for the activating effects and cell permeability of CZ-48 were identified. CZ-48 activated SARM1 via a conformational change of the auto-inhibitory domain and dimerization of its catalytic domain. SARM1 catalysis was similar to CD38, despite having no sequence similarity. Both catalyzed similar set of reactions, but SARM1 had much higher NAD-cyclizing activity, making it more efficient in elevating cADPR. CZ-48 acted selectively, activating SARM1 but inhibiting CD38. In SARM1-overexpressing cells, CZ-48 elevated cADPR, depleted NAD and ATP, and induced non-apoptotic death. CZ-48 is a specific modulator of SARM1 functions in cells. : Biochemistry; Enzymology; Biochemical Mechanism Subject Areas: Biochemistry, Enzymology, Biochemical Mechanism
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- 2019
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6. Direct Gating of the TRPM2 Channel by cADPR via Specific Interactions with the ADPR Binding Pocket
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Peilin Yu, Zhenming Liu, Xiafei Yu, Peiwu Ye, Huan Liu, Xiwen Xue, Lixin Yang, Zhongtang Li, Yang Wu, Cheng Fang, Yong Juan Zhao, Fan Yang, Jian Hong Luo, Lin-Hua Jiang, Liangren Zhang, Lihe Zhang, and Wei Yang
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Biology (General) ,QH301-705.5 - Abstract
Summary: cADPR is a well-recognized signaling molecule by modulating the RyRs, but considerable debate exists regarding whether cADPR can bind to and gate the TRPM2 channel, which mediates oxidative stress signaling in diverse physiological and pathological processes. Here, we show that purified cADPR evoked TRPM2 channel currents in both whole-cell and cell-free single-channel recordings and specific binding of cADPR to the purified NUDT9-H domain of TRPM2 by surface plasmon resonance. Furthermore, by combining computational modeling with electrophysiological recordings, we show that the TRPM2 channels carrying point mutations at H1346, T1347, L1379, S1391, E1409, and L1484 possess distinct sensitivity profiles for ADPR and cADPR. These results clearly indicate cADPR is a bona fide activator at the TRPM2 channel and clearly delineate the structural basis for cADPR binding, which not only lead to a better understanding in the gating mechanism of TRPM2 channel but also shed light on a cADPR-induced RyRs-independent Ca2+ signaling mechanism. : Yu et al. demonstrate that cADPR, which was thought to be an intracellular calcium messenger by modulating function of the ryanodine receptors (RyRs), also activates the TRPM2 channel. Their results suggest that cADPR regulates intracellular calcium release through multiple pathways. Keywords: cADPR, NUDT9-H domain, TRPM2 channel
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- 2019
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7. Cyclic ADP-ribose and heat regulate oxytocin release via CD38 and TRPM2 in the hypothalamus during social or psychological stress in mice
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Jing Zhong, Sarwat Amina, Mingkun Liang, Shirin Akther, Teruko Yuhi, Tomoko Nishimura, Chiharu Higashida, Takahiro Tsuji, Hong-Xiang Liu, Minako Hashii, Kazumi Furuhara, Shigeru Yokoyama, Yasuhiko Yamamoto, Hiroshi Okamoto, Yong Juan Zhao, Hon Cheung Lee, Makoto Tominaga, Olga Lopatina, and Haruhiro Higashida
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Cyclic ADP-Ribose ,Oxytocin ,stress ,hyperthermia ,secretion ,Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Hypothalamic oxytocin (OT) is released into the brain by cyclic ADP-ribose (cADPR) with or without depolarizing stimulation. Previously, we showed that the intracellular free calcium concentration ([Ca2+]i) that seems to trigger OT release can be elevated by -NAD+, cADPR, and ADP in mouse oxytocinergic neurons. As these -NAD+ metabolites activate warm-sensitive TRPM2 cation channels, when the incubation temperature is increased, the [Ca2+]i in hypothalamic neurons is elevated. However, it has not been determined whether OT release is facilitated by heat in vitro or hyperthermia in vivo in combination with cADPR. Furthermore, it has not been examined whether CD38 and TRPM2 exert their functions on OT release during stress or stress-induced hyperthermia in relation to the anxiolytic roles and social behaviors of OT under stress conditions. Here, we report that OT release from the isolated hypothalami of male mice in culture was enhanced by extracellular application of cADPR or increasing the incubation temperature from 35°C to 38.5°C, and simultaneous stimulation showed a greater effect. This release was inhibited by a cADPR-dependent ryanodine receptor inhibitor and a nonspecific TRPM2 inhibitor. The facilitated release by heat and cADPR was suppressed in the hypothalamus isolated from CD38 knockout mice and CD38- or TRPM2-knockdown mice. In the course of these experiments, we noted that OT release differed markedly between individual mice under stress with group housing. That is, when male mice received cage-switch stress and eliminated due to their social subclass, significantly higher levels of OT release were found in subordinates compared with ordinates. In mice exposed to anxiety stress in an open field, the cerebrospinal fluid (CSF) OT level increased transiently at 5 minutes after exposure, and the rectal temperature also increased from 36.6°C to 37.8°C. OT levels in the CSF of mice with lipopolysaccharide-induced fever (+0.8°C) were higher than those of control mice. The TRPM2 mRNA levels and immunoreactivities increased in the subordinate group with cage-switch stress. These results showed that cADPR/CD38 and heat/TRPM2 are co-regulators of OT secretion and suggested that CD38 and TRPM2 are potential therapeutic targets for OT release in psychiatric diseases caused by social stress.
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- 2016
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8. Role of EtMIC4 EGF-like in regulating the apoptosis of Eimeria tenella host cells via the EGFR pathway
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Xue-song Zhang, Yong-juan Zhao, Yu Zhang, Tong Xu, Kai-ling Cui, Bu-ting Duan, Xiao-ling Lv, Li Zhang, Zhi-yong Xu, Rui Bai, and Ming-xue Zheng
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Eimeria tenella ,EtMIC4 EGF-like ,apoptosis ,EGFR pathway ,Animal culture ,SF1-1100 - Abstract
This study aimed to explore the role and key point of EtMIC4 EGF-like recombinant protein in regulating the apoptosis of Eimeria tenella host cells via the epidermal growth factor receptor (EGFR) pathway. The cells were treated with EtMIC4 EGF-like protein, EGFR-specific siRNA, or both. Infection and apoptosis rates as well as dynamic changes in the key genes and proteins of the EGFR signaling pathway in the host cells were determined. Results showed that the E. tenella and EtMIC4 EGF-like group had the highest infection rate (P < 0.01). In cells treated with EtMIC4 EGF-like for 4 to 24 h, the apoptosis rate was significantly decreased (P < 0.01) and the relative mRNA expression and protein phosphorylation levels of EGFR, protein kinase B (AKT), and extracellular regulated protein kinases (ERK) were significantly increased (P < 0.01). In E. tenella sporozoites infected for 4 to 96 h, the rate of host cell apoptosis induced by E. tenella infection was significantly (P < 0.01) reduced by EtMIC4 EGF-like. The relative mRNA expression and protein phosphorylation levels of EGFR, AKT, and ERK in the host cells of E. tenella + EtMIC4 EGF-like group were significantly increased (P < 0.01). These results indicated that E. tenella could activate the EGFR pathway through EtMIC4 EGF-like and regulate the expression of key genes in the AKT and ERK signaling pathways, thereby inhibiting cell apoptosis.
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- 2022
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9. Base-Exchange Enabling the Visualization of SARM1 Activities in Sciatic Nerve-Injured Mice
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Ke Huang, Wen Jie Zhu, Wan Hua Li, Hon Cheung Lee, Yong Juan Zhao, and Chi-Sing Lee
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Fluid Flow and Transfer Processes ,Process Chemistry and Technology ,Bioengineering ,Instrumentation - Published
- 2023
10. ENZYMATIC AND ENDOGENOUS SYNTHESIS OF NAD(P)‐DERIVED CALCIUM‐MOBILIZING MESSENGERS
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Hon Cheung Lee and Yong Juan Zhao
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- 2022
11. Establishment of an efficient regeneration and genetic transformation system for Malus prunifolia Borkh. ‘Fupingqiuzi’
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Yu-song LIU, Hong-ying WANG, Yong-juan ZHAO, Yi-bo JIN, Chao LI, and Feng-wang MA
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Food Animals ,Ecology ,Animal Science and Zoology ,Plant Science ,Agronomy and Crop Science ,Biochemistry ,Food Science - Published
- 2022
12. [Application of high content imaging technique in research on effective components of traditional Chinese medicine: a review]
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Yong-Juan, Zhao, Wen, Gao, Xu-Dong, Xing, and Hua, Yang
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Medicine, Chinese Traditional ,Drugs, Chinese Herbal - Abstract
High content imaging(HCI) technique that combines automatic high throughput with high-resolution cell imaging, is characterized by abundant data information, high imaging sensitivity, easy visualization and standardization, and is commonly used in the cellular(or subcellular) phenotypic analysis. Abundant phenotypic information can be obtained by using HCI in one experiment, including cellular morphology, cellular structure, and signal transduction pathways of related functions, on the basis of the maintenance of the integrity of cellular structures and functions. Multiple studies have shown that a series of dynamic spatio-temporal interactive change processes were induced by the disturbance of cells by specific factors, making cell phenotypes change accordingly, especially for the slight perturbation response of cells. Generally, the detection of one or several endpoint effect indicators is often difficult to accurately and comprehensively reflect the overall efficacy information of traditional Chinese medicine(TCM) because of its unique characteristics of multi-components and multi-targets. The application of HCI is thus helpful to discover the effective components and their action modes in the complex system of TCM. This paper reviewed the application progress in the HCI technique in the screening of active components and their regulation mechanism to provide references for further research.
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- 2022
13. Acidic pH irreversibly activates the signaling enzyme SARM1
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Dapeng Zhang, Zhi Ying Zhao, Wei Ming He, Yongjun Tan, Wan Hua Li, Yun Nan Hou, Yong Juan Zhao, and Qian Wen Wang
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Models, Molecular ,Niacinamide ,medicine.disease_cause ,Biochemistry ,chemistry.chemical_compound ,Protein Domains ,Chaps ,Disulfiram ,medicine ,Humans ,Enzyme Inhibitors ,Molecular Biology ,Nicotinamide mononucleotide ,Armadillo Domain Proteins ,chemistry.chemical_classification ,Mutation ,Nicotinamide ,Chemistry ,Mutagenesis ,Cell Biology ,Hydrogen-Ion Concentration ,NAD ,Enzyme Activation ,Cytoskeletal Proteins ,Kinetics ,HEK293 Cells ,Enzyme ,Biocatalysis ,Biophysics ,NAD+ kinase ,Salt bridge ,Acids - Abstract
SARM1, an executioner in axon degeneration, is an autoinhibitory NAD-consuming enzyme, composed of multiple domains. NMN and its analogs, CZ-48 and VMN, are the only known activators, which can release the inhibitory ARM domain from the enzymatic TIR domain. Here, we document that acid can also activate SARM1, even more efficiently than NMN, possibly via the protonation of the negative residues. Systematic mutagenesis revealed that a single mutation, E689Q in TIR, led to the constitutive activation of SARM1. It forms a salt bridge with R216 in the neighboring ARM, maintaining the autoinhibitory structure. Using this 'acid activation' protocol, mutation K597E was found to inhibit activation, while H685A eliminated SARM1 catalytic activity, revealing two distinct inhibitory mechanisms. The protocol has also been applied to differentiate two classes of chemical inhibitors. NAD, dHNN, disulfiram, CHAPS, and TRX-100 mainly inhibited the activation process, while nicotinamide and Tweens mainly inhibited SARM1 catalysis. Taken together, we demonstrate a new mechanism for SARM1 activation and decipher two distinct inhibitory mechanisms of SARM1.
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- 2021
14. Vertebra segmentation of spine MRI with improved GVF snake based on shape knowledge.
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Yong-Juan Zhao, Lin Shi 0001, Jia-Chun Li, James F. Griffith, Anil T. Ahuja, and Pheng-Ann Heng
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- 2011
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15. GALA peptide improves the potency of nanobody–drug conjugates by lipid-induced helix formation
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Zhengshuang Xu, Xiao Chun Guo, Yong Juan Zhao, Hon Cheung Lee, Ya Jie Chen, Jian Yuan Yang, Qi Wen Deng, Li Wang, and Ting Li
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Antineoplastic Agents ,Peptide ,Nanoconjugates ,010402 general chemistry ,Endocytosis ,01 natural sciences ,Catalysis ,law.invention ,law ,Cell Line, Tumor ,Amphiphile ,Materials Chemistry ,Humans ,Cytotoxicity ,chemistry.chemical_classification ,010405 organic chemistry ,Chemistry ,Metals and Alloys ,General Chemistry ,Recombinant Proteins ,0104 chemical sciences ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Helix ,Ceramics and Composites ,Biophysics ,Recombinant DNA ,Peptides ,Oligopeptides ,Alpha helix ,Single-Chain Antibodies ,Conjugate - Abstract
A novel nanobody-drug conjugate (NDC) was constructed by incorporating an amphipathic peptide, GALA, which improved the cytotoxicity by one to two orders of magnitude. Mechanistic studies demonstrate that tethering to lipids induces GALA to form a helix, which dramatically enhances endocytosis. Our work provides a general strategy not only for improving the anti-cancer efficacy of protein-drug conjugates but also for increasing the efficiency of other types of endocytosis-dependent cell delivery.
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- 2021
16. The complex structure of GRL0617 and SARS-CoV-2 PLpro reveals a hot spot for antiviral drug discovery
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Dan Cao, Ziyang Fu, Jinle Tang, Yuxian Xiong, Yong Juan Zhao, Zhihong Liu, Guoliang Zhang, Jihui Li, Hao Huang, Huan Zhou, Ming Liu, Wenning Zhu, Bin Huang, Shuyan Liu, Yuxin Ye, and Xiaogang Niu
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0301 basic medicine ,Models, Molecular ,medicine.drug_class ,Protein Conformation ,medicine.medical_treatment ,viruses ,Science ,General Physics and Astronomy ,01 natural sciences ,Antiviral Agents ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Inhibitory Concentration 50 ,medicine ,Virus maturation ,Humans ,Drug Interactions ,skin and connective tissue diseases ,Pandemics ,Ubiquitins ,Coronavirus 3C Proteases ,Multidisciplinary ,Protease ,010405 organic chemistry ,Chemistry ,Drug discovery ,SARS-CoV-2 ,Nanocrystallography ,HEK 293 cells ,fungi ,COVID-19 ,General Chemistry ,Cysteine protease ,ISG15 ,0104 chemical sciences ,Cell biology ,High-Throughput Screening Assays ,COVID-19 Drug Treatment ,body regions ,030104 developmental biology ,HEK293 Cells ,Structural biology ,Viral infection ,Cytokines ,Antiviral drug - Abstract
SARS-CoV-2 is the pathogen responsible for the COVID-19 pandemic. The SARS-CoV-2 papain-like cysteine protease (PLpro) has been implicated in playing important roles in virus maturation, dysregulation of host inflammation, and antiviral immune responses. The multiple functions of PLpro render it a promising drug target. Therefore, we screened a library of approved drugs and also examined available inhibitors against PLpro. Inhibitor GRL0617 showed a promising in vitro IC50 of 2.1 μM and an effective antiviral inhibition in cell-based assays. The co-crystal structure of SARS-CoV-2 PLproC111S in complex with GRL0617 indicates that GRL0617 is a non-covalent inhibitor and it resides in the ubiquitin-specific proteases (USP) domain of PLpro. NMR data indicate that GRL0617 blocks the binding of ISG15 C-terminus to PLpro. Using truncated ISG15 mutants, we show that the C-terminus of ISG15 plays a dominant role in binding PLpro. Structural analysis reveals that the ISG15 C-terminus binding pocket in PLpro contributes a disproportionately large portion of binding energy, thus this pocket is a hot spot for antiviral drug discovery targeting PLpro., The SARS-CoV-2 papain-like protease (PLpro) is of interest as a drug target. Here, the authors identify GRL0617 as a PPI (protein–protein interaction) inhibitor of SARS-CoV-2 PLpro that inhibits its deISGylating activity and present the mechanism of action of the compound through the GRL0617-bound PLpro crystal structure and NMR studies.
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- 2021
17. The mitigation effects of exogenous dopamine on low nitrogen stress in Malus hupehensis
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Zhijun Zhang, Ma FengWang, Yi-bo Jin, Xiaomin Liu, Kexin Tan, Tengteng Gao, Yong-juan Zhao, and Chao Li
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0106 biological sciences ,Agriculture (General) ,Plant Science ,Nitrate reductase ,01 natural sciences ,Biochemistry ,S1-972 ,chemistry.chemical_compound ,Food Animals ,nitrogen deficiency ,Dopamine ,Glutamine synthetase ,ethylene ,medicine ,Malus hupehensis ,root system architecture ,Ecology ,biology ,04 agricultural and veterinary sciences ,Glutamic acid ,Metabolism ,Nitrite reductase ,biology.organism_classification ,chemistry ,Chlorophyll ,040103 agronomy & agriculture ,0401 agriculture, forestry, and fisheries ,Animal Science and Zoology ,dopamine ,Agronomy and Crop Science ,010606 plant biology & botany ,Food Science ,medicine.drug - Abstract
Dopamine plays numerous physiological roles in plants. We explored its role in the regulation of growth, nutrient absorption, and response to nitrogen (N) deficiency in Malus hupehensis Rehd. Under low N condition, plant growth slowed, and the net photosynthetic rates, chlorophyll contents, and maximal quantum yield of PSII (Fv/Fm) decreased significantly. However, the application of 100 μmol L−1 exogenous dopamine significantly reduced the inhibition of low N stress on plant growth. In addition to modifying root system architecture under low N supply, exogenous dopamine also changed the uptake, transport, and distribution of N, P, and K. Furthermore, exogenous dopamine enhances the tolerance to low nitrogen stress by increasing the activity of enzymes (nitrate reductase, nitrite reductase, glutamic acid synthase and glutamine synthetase) involved in N metabolism. We also found that exogenous dopamine promoted the expression of ethylene signaling genes (ERF1, ERF2, EIL1, ERS2, ETR1, and EIN4) under low N stress. Therefore, we hypothesized that ethylene might be involved in dopamine response to low N stress in M. hupehensis. Our results suggest that exogenous dopamine can mitigate low N stress by regulating the absorption of mineral nutrients, possibly through the regulation of the ethylene signaling pathway.
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- 2020
18. The role of Ca
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Xiao-Ling, Lv, Yong-Yan, Wang, Ming-Xue, Zheng, Rui, Bai, Li, Zhang, Bu-Ting, Duan, Xuan, Lei, Xue-Song, Zhang, Yong-Juan, Zhao, Kai-Ling, Cui, and Tong, Xu
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Coccidiosis ,Animals ,Chick Embryo ,Cecum ,Chickens ,Eimeria tenella ,Poultry Diseases - Abstract
Cecal epithelial cell damage is a key factor in host injure during the development of E. tenella. The intracellular free Ca
- Published
- 2022
19. Resolving the topological enigma in Ca2+ signaling by cyclic ADP-ribose and NAADP
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Yong Juan Zhao and Hon Cheung Lee
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0301 basic medicine ,Nicotinic acid adenine dinucleotide phosphate ,030102 biochemistry & molecular biology ,Endoplasmic reticulum ,Cell Biology ,Topology ,Biochemistry ,Cyclic ADP-ribose ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,chemistry ,NAD+ kinase ,Signal transduction ,Molecular Biology ,Biogenesis ,Calcium signaling ,Nicotinamide mononucleotide - Abstract
Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) are two structurally distinct messengers that mobilize the endoplasmic and endolysosomal Ca2+ stores, respectively. Both are synthesized by the CD38 molecule (CD38), which has long been thought to be a type II membrane protein whose catalytic domain, intriguingly, faces to the outside of the cell. Accordingly, for more than 20 years, it has remained unresolved how CD38 can use cytosolic substrates such as NAD and NADP to produce messengers that target intracellular Ca2+ stores. The discovery of type III CD38, whose catalytic domain faces the cytosol, has now begun to clarify this topological conundrum. This article reviews the ideas and clues leading to the discovery of the type III CD38; highlights an innovative approach for uncovering its natural existence; and discusses the regulators of its activity, folding, and degradation. We also review the compartmentalization of cADPR and NAADP biogenesis. We further discuss the possible mechanisms that promote type III CD38 expression and appraise a proposal of a Ca2+-signaling mechanism based on substrate limitation and product translocation. The surprising finding of another enzyme that produces cADPR and NAADP, sterile α and TIR motif–containing 1 (SARM1), is described. SARM1 regulates axonal degeneration and has no sequence similarity with CD38 but can catalyze the same set of multireactions and has the same cytosolic orientation as the type III CD38. The intriguing finding that SARM1 is activated by nicotinamide mononucleotide to produce cADPR and NAADP suggests that it may function as a regulated Ca2+-signaling enzyme like CD38.
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- 2019
20. The transferrin receptor CD71 regulates type II CD38, revealing tight topological compartmentalization of intracellular cyclic ADP-ribose production
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Lei Fang, Qi Wen Deng, Hon Cheung Lee, Wei Ming He, Ting Li, Yong Juan Zhao, and Jingzi Zhang
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0301 basic medicine ,Cytoplasm ,Transferrin receptor ,CD38 ,Biochemistry ,Cyclic ADP-ribose ,03 medical and health sciences ,chemistry.chemical_compound ,Cytosol ,Antigens, CD ,immune system diseases ,hemic and lymphatic diseases ,Receptors, Transferrin ,Humans ,Molecular Biology ,Cyclic ADP-Ribose ,030102 biochemistry & molecular biology ,Endoplasmic reticulum ,Cell Membrane ,hemic and immune systems ,Cell Biology ,ADP-ribosyl Cyclase 1 ,Cell biology ,HEK293 Cells ,030104 developmental biology ,chemistry ,Membrane topology ,Calcium ,Biogenesis ,Intracellular ,Signal Transduction ,Protein Binding - Abstract
The CD38 molecule (CD38) catalyzes biogenesis of the calcium-mobilizing messenger cyclic ADP-ribose (cADPR). CD38 has dual membrane orientations, and type III CD38, with its catalytic domain facing the cytosol, has low abundance but is efficient in cyclizing cytosolic NAD to produce cADPR. The role of cell surface type II CD38 in cellular cADPR production is unknown. Here we modulated type II CD38 expression and assessed the effects of this modulation on cADPR levels. We developed a photoactivatable cross-linking probe based on a CD38 nanobody, and, combining it with MS analysis, we discovered that cell surface CD38 interacts with CD71. CD71 knockdown increased CD38 levels, and CD38 knockout reciprocally increased CD71, and both could be cocapped and coimmunoprecipitated. We constructed a chimera comprising the N-terminal segment of CD71 and a CD38 nanobody to mimic CD71's ligand property. Overexpression of this chimera induced a dramatically large decrease in CD38 via lysosomes. Remarkably, cellular cADPR levels did not decrease correspondingly. Bafilomycin-mediated blockade of lysosomal degradation greatly elevated active type II CD38 by trapping it in the lysosomes but also did not increase cADPR levels. Retention of type II CD38 in the endoplasmic reticulum (ER) by expressing an ER construct that prevented its transport to the cell surface likewise did not change cADPR levels. These results provide first and direct evidence that cADPR biogenesis occurs in the cytosol and is catalyzed mainly by type III CD38 and that type II CD38, compartmentalized in the ER or lysosomes or on the cell surface, contributes only minimally to cADPR biogenesis.
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- 2019
21. A non-internalised CD38-binding radiolabelled single-domain antibody fragment to monitor and treat multiple myeloma
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Yoline Lauwers, Yves Beguin, Mireille Dumoulin, Jo Caers, Margaux Lejeune, Frédéric Baron, Ahmet Krasniqi, Matthias D'Huyvetter, Yong Juan Zhao, Nick Devoogdt, Elodie Duray, Supporting clinical sciences, Medical Imaging, and Faculty of Medicine and Pharmacy
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Cancer Research ,medicine.medical_specialty ,Biodistribution ,Single Photon Emission Computed Tomography Computed Tomography ,medicine.drug_class ,Pharmacology ,Lutetium ,Monoclonal antibody ,Mice ,Radio-immunotherapy ,In vivo ,Internal medicine ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Diseases of the blood and blood-forming organs ,Tissue Distribution ,Molecular Biology ,Multiple myeloma ,RC254-282 ,Camelidae ,single-domain antibody ,Radioisotopes ,Hematology ,biology ,business.industry ,Research ,Daratumumab ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Single-Domain Antibodies ,medicine.disease ,ADP-ribosyl Cyclase 1 ,multiple myeloma ,Single-domain antibody ,Oncology ,Theranostic ,Radiology Nuclear Medicine and imaging ,biology.protein ,Nanobody ,Antibody ,RC633-647.5 ,TARGETED RADIONUCLIDE THERAPY ,business ,CD38 - Abstract
Background Antibody-based therapies targeting CD38 are currently used as single agents as well as in combination regimens for multiple myeloma, a malignant plasma cell disorder. In this study, we aimed to develop anti-CD38 single-domain antibodies (sdAbs) that can be used to trace CD38+ tumour cells and subsequently used for targeted radionuclide therapy. SdAbs are derived from Camelidae heavy-chain antibodies and have emerged as promising theranostic agents due to their favourable pharmacological properties. Methods Four different anti-CD38 sdAbs were produced, and their binding affinities and potential competition with the monoclonal antibody daratumumab were tested using biolayer interferometry. Their binding kinetics and potential cell internalisation were further studied after radiolabelling with the diagnostic radioisotope Indium-111. The resulting radiotracers were evaluated in vivo for their tumour-targeting potential and biodistribution through single-photon emission computed tomography (SPECT/CT) imaging and serial dissections. Finally, therapeutic efficacy of a lead anti-CD38 sdAb, radiolabelled with the therapeutic radioisotope Lutetium-177, was evaluated in a CD38+ MM xenograft model. Results We retained anti-CD38 sdAb #2F8 as lead based on its excellent affinity and superior stability, the absence of competition with daratumumab and the lack of receptor-mediated internalisation. When intravenously administered to tumour-xenografted mice, radiolabelled sdAb #2F8 revealed specific and sustained tumour retention with low accumulation in other tissues, except kidneys, resulting in high tumour-to-normal tissue ratios. In a therapeutic setting, myeloma-bearing mice received three consecutive intravenous administrations of a high (18.5 MBq) or a low radioactive dose (9.3 MBq) of 177Lu-DTPA-2F8 or an equal volume of vehicle solution. A dose-dependent tumour regression was observed, which translated into a prolonged median survival from 43 days for vehicle-treated mice, to 62 days (p = 0.027) in mice receiving the low and 65 days in mice receiving the high (p = 0.0007) radioactive dose regimen, respectively. Conclusions These results highlight the theranostic potential of radiolabelled anti-CD38 sdAbs for the monitoring and treatment of multiple myeloma.
- Published
- 2021
22. Author response: Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate
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Ke Huang, Wan Hua Li, Hon Cheung Lee, Hongmin Zhang, Yang Cai, Yun Nan Hou, Sujing Wang, Sheng Cao, Qian Wen Wang, Yang Du, Xu Jie Xie, Yong Juan Zhao, Chi-Sing Lee, Wen Jie Zhu, and Zhi Ying Zhao
- Subjects
Drug candidate ,Chemistry ,Fluorescence ,Cell biology - Published
- 2021
23. Rational Design and Identification of Small‐Molecule Allosteric Inhibitors of CD38
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Lihe Zhang, Songlu Li, Hon Cheung Lee, Xiaomeng Shi, Yong Juan Zhao, Hongwei Jin, Zhenming Liu, Liangren Zhang, Ting Li, Lixin Yang, and Yang Wu
- Subjects
Protein Conformation ,Allosteric regulation ,Molecular Dynamics Simulation ,Nicotinamide adenine dinucleotide ,Antibodies, Monoclonal, Humanized ,010402 general chemistry ,01 natural sciences ,Biochemistry ,chemistry.chemical_compound ,Allosteric Regulation ,Humans ,Enzyme Inhibitors ,Molecular Biology ,Membrane Glycoproteins ,Nicotinic acid adenine dinucleotide phosphate ,010405 organic chemistry ,Chemistry ,Drug discovery ,Organic Chemistry ,Rational design ,Ligand (biochemistry) ,ADP-ribosyl Cyclase 1 ,Small molecule ,0104 chemical sciences ,Drug Design ,Biophysics ,Molecular Medicine ,NAD+ kinase - Abstract
CD38 is a multi-functional signaling enzyme that catalyzes the biosynthesis of two calcium-mobilizing second messengers: cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate. It also regulates intracellular nicotinamide adenine dinucleotide (NAD) contents, associated with multiple pathophysiological processes such as aging and cancer. As such, enzymatic inhibitors of CD38 offer great potential in drug development. Here, through virtual screening and enzymatic assays, we discovered compound LX-102, which targets CD38 on the side opposite its enzymatic pocket with a binding affinity of 7.7 μm. It inhibits the NADase activity of CD38 with an IC50 of 14.9 μm. Surface plasmon resonance (SPR) and hydrogen/deuterium exchange and mass spectrometry experiments verified that LX-102 competitively binds to the epitope of the therapeutic SAR 650984 antibody in an allosteric manner. Molecular dynamics simulation was performed to demonstrate the binding dynamics of CD38 with the allosteric ligand. In summary, we established that the cavity to which SAR 650984 binds was an allosteric site and was accessible for the rational design of small chemical modulators of CD38. The lead compound LX-102 that we identified in this study could also be a useful tool for probing CD38 functions and promoting drug discovery.
- Published
- 2019
24. A cytosolic chaperone complex controls folding and degradation of type III CD38
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Yong Juan Zhao, Yang Wu, Lei Fang, Jingzi Zhang, and Hon Cheung Lee
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0301 basic medicine ,Protein Folding ,Immunoprecipitation ,Protein degradation ,Biochemistry ,03 medical and health sciences ,Cytosol ,immune system diseases ,hemic and lymphatic diseases ,Humans ,Molecular Biology ,Gene knockdown ,Membrane Glycoproteins ,030102 biochemistry & molecular biology ,biology ,Chemistry ,hemic and immune systems ,Cell Biology ,ADP-ribosyl Cyclase 1 ,Hsp90 ,Transmembrane protein ,Cell biology ,HEK293 Cells ,030104 developmental biology ,Protein Synthesis and Degradation ,Chaperone (protein) ,biology.protein ,Chaperone complex ,Protein folding - Abstract
Cluster of differentiation 38 (CD38) is the best-studied enzyme catalyzing the synthesis of the Ca(2+) messenger cyclic ADP-ribose. It is a single-pass transmembrane protein, but possesses dual orientations. We have documented the natural existence of type III CD38 in cells and shown that it is regulated by a cytosolic activator, calcium- and integrin-binding 1 (CIB1). However, how type III CD38 can be folded correctly in the reductive cytosol has not been addressed. Using the yeast two-hybrid technique with CD38's catalytic domain (sCD38) as bait, here we identified a chaperone, Hsp70-interacting protein (Hip), that specifically interacts with both the type III CD38 and sCD38. Immunoprecipitation coupled with MS identified a chaperone complex associated specifically with sCD38. Pharmacological and siRNA-mediated knockdown of Hsp90 chaperones decreased the expression levels of both sCD38 and type III CD38, suggesting that these chaperones facilitate their folding. Moreover, knockdown of Hsc70 or DNAJA2 increased the levels of both CD38 types, consistent with the roles of these proteins in mediating CD38 degradation. Notably, Hip knockdown decreased type III CD38 substantially, but only marginally affected sCD38, indicating that Hip was selective for the former. More remarkably, DNAJA1 knockdown decreased sCD38 but increased type III CD38 levels. Mechanistically, we show that Hsc70 mediates lysosomal degradation of type III CD38, requiring the lysosomal receptor Lamp2A and the C19-motif in the C terminus of CD38. Our results indicate that folding and degradation of type III CD38 is effectively controlled in cells, providing further strong support of its physiological relevance.
- Published
- 2019
25. Anti-Multiple Myeloma Activity of Nanobody-Based Anti-CD38 Chimeric Antigen Receptor T Cells
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Huan Chen, Hon Cheung Lee, Yun Nan Hou, Na An, Ting Li, Cheng Fang, Qiong Li Zhang, Qiao Xia Zhang, Yong Juan Zhao, and Xin Du
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0301 basic medicine ,CD3 ,Receptors, Antigen, T-Cell ,CD34 ,Pharmaceutical Science ,Mice, SCID ,CD38 ,Flow cytometry ,Interferon-gamma ,Mice ,03 medical and health sciences ,0302 clinical medicine ,immune system diseases ,hemic and lymphatic diseases ,Drug Discovery ,medicine ,Animals ,Humans ,Cytotoxicity ,Receptors, Chimeric Antigen ,medicine.diagnostic_test ,biology ,Tumor Necrosis Factor-alpha ,Chemistry ,hemic and immune systems ,Single-Domain Antibodies ,ADP-ribosyl Cyclase 1 ,Chimeric antigen receptor ,030104 developmental biology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Interleukin-2 ,Molecular Medicine ,K562 Cells ,Multiple Myeloma ,K562 cells - Abstract
Chimeric antigen receptor T cells (CAR-Ts) are a promising strategy for the treatment of many cancers, including multiple myeloma (MM), a hematological malignancy characterized by the high expression of CD38. To broaden the applications of using CD38 as a therapeutic target for the disease, we developed a new nanobody against CD38 and constructed a CD38-CAR that was composed of this nanobody as the targeting domain, and 4-1BB and CD3ζ as the costimulatory and activating domains, in a lentiviral vector. CD3+ T cells from healthy individuals were transduced with the CD38-CAR at an efficiency higher than 60%, as determined by CD38-CAR expression using flow cytometry. The CD38-CAR-Ts proliferated efficiently and produced more inflammatory cytokines, such as IL-2, IFN-γ, and TNF-α, when activated. The CD38-CAR-Ts effectively lysed CD38+ MM cell lines, including LP-1, RPMI 8226, OPM2, and MOLP8, and primary MM cells from multiple myeloma patients. The specificity was demonstrated by the fact that CD38-CAR-Ts showed little cytotoxicity on LP-1 cells with CD38 knocked out or on K562 cells, which do not express CD38. CD38-CAR-Ts appeared to have a very slight cytotoxicity against CD38+ fractions of T cells, B cells, and natural killer cells. In addition, the lysis of CD34+ hematopoietic progenitor cells did not completely inhibit the development of colony-forming units. In vivo, CD38-CAR-Ts inhibited tumor growth in NOD/SCID mice that were subcutaneously inoculated with RPMI 8226 cells. These results demonstrate that the CD38-CAR-Ts constructed with the anti-CD38 nanobody are a promising approach for the treatment of multiple myeloma.
- Published
- 2018
26. Permeant Fluorescent Probes Visualize the Activation of SARM1 and Uncover an Anti-neurodegenerative Drug Candidate
- Author
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Yun Nan Hou, Yang Du, Ke Huang, Sheng Cao, Xu Jie Xie, Hon Cheung Lee, Yang Cai, Hongmin Zhang, Wen Jie Zhu, Chi-Sing Lee, Sujing Wang, Zhi Ying Zhao, Yong Juan Zhao, Wan Hua Li, and Qian Wen Wang
- Subjects
Chemistry ,Drug candidate ,Covalent bond ,medicine ,Biophysics ,Nisoldipine ,Endogeny ,Degeneration (medical) ,Fluorescence ,Axonal degeneration ,Neuroprotection ,medicine.drug - Abstract
SARM1 regulates axonal degeneration through its NAD-metabolizing activity and is a drug target for neurodegenerative disorders. We designed and synthesized fluorescent conjugates of styryl derivative with pyridine to serve as substrates of SARM1, which exhibited large red-shifts after conversion. With the conjugates, SARM1 activation was visualized in live cells following elevation of endogenous NMN or treatment with a cell-permeant NMN-analog. In neurons, imaging documented SARM1 activation preceded vincristine-induced axonal degeneration by hours. Library screening identified a derivative of nisoldipine as a covalent inhibitor of SARM1 that reacted with Cys311 in its Armadillo-domain and blocked its NMN-activation, protecting axons from degeneration. CryoEM showed that SARM1 was locked into an inactive conformation by the inhibitor, uncovering an unsuspected neuroprotective mechanism of dihydropyridines.
- Published
- 2021
27. ImmunoPET Imaging of Multiple Myeloma with [68Ga]Ga-NOTA-Nb1053
- Author
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Cheng Wang, Yumei Chen, Yun Nan Hou, Qiufang Liu, Di Zhang, Haitao Zhao, You Zhang, Shuxian An, Lianghua Li, Jian Hou, Gang Huang, Jianjun Liu, Yong Juan Zhao, and Weijun Wei
- Abstract
Purpose Multiple myeloma (MM) remains incurable and its diagnosis relies heavily on bone marrow aspiration and biopsy. CD38 is a glycoprotein highly specific for MM. Antibody therapeutics (e.g., daratumumab) targeting CD38 have shown encouraging efficacy in treating MM, either as a monotherapy agent or in combination with other regimens. However, efficient stratification of patients who might benefit from daratumumab therapy and timely monitoring of the therapeutic responses are still clinical challenges. This work aims to devise a CD38-targeted imaging strategy and assess its value in diagnosing MMs. Methods By labeling a CD38-specific single domain antibody (Nb1053) with 68Ga (t1/2 = 1.1 h), we developed a CD38-targeted immuno-positron emission tomography (immunoPET) imaging probe [68Ga]Ga-NOTA-Nb1053. The probe was developed with good radiochemical yield (> 50%), excellent radiochemical purity (> 99%), and immunoreactivity (> 95%). The diagnostic accuracy of the probe was thoroughly investigated in preclinical MM models. Results ImmunoPET imaging with the probe specifically depicted all the subcutaneous and orthotopic MM lesions, outperforming the traditional 18F-fluorodeoxyglucose PET and the nonspecific [68Ga]Ga-NOTA-NbGFP immunoPET. More importantly, daratumumab preloading significantly reduced [68Ga]Ga-NOTA-Nb1053 uptake in the disseminated bone lesions, indicating the overlapping targeting epitopes of [68Ga]Ga-NOTA-Nb1053 with that of daratumumab. Furthermore, premedication with sodium maleate or fructose significantly decreased kidney retention of [68Ga]Ga-NOTA-Nb1053 and improved the diagnostic value of the probe in lymphoma models. Conclusion This work successfully developed a novel CD38-targeted immunoPET imaging approach that enabled precise visualization of CD38 and diagnosis of MMs. Upon clinical translation, [68Ga]Ga-NOTA-Nb1053 immunoPET may serve as a valuable CD38-targeted molecular imaging toolbox, facilitating early diagnosis of MM and precise assessment of the therapeutic responses.
- Published
- 2021
28. MdTyDc Overexpression Improves Alkalinity Tolerance in Malus domestica
- Author
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Yi-bo Jin, Chao Li, Xiaomin Liu, Kexin Tan, Tengteng Gao, Fengwang Ma, Jiangzhu Zheng, Yong-juan Zhao, and Zhijun Zhang
- Subjects
Malus ,MdTyDc ,biology ,Membrane permeability ,Chemistry ,Abiotic stress ,fungi ,Plant Science ,lcsh:Plant culture ,Tyramine ,biology.organism_classification ,Photosynthetic capacity ,Tyrosine decarboxylase ,chemistry.chemical_compound ,Ion homeostasis ,Biochemistry ,N metabolism ,lcsh:SB1-1110 ,ion homeostasis ,Tyrosine ,alkaline stress ,dopamine ,Original Research - Abstract
Tyrosine is decarboxylated to tyramine by TYDC (Tyrosine decarboxylase) and then hydroxylated to dopamine, which is involved in plant response to abiotic stress. However, little is known about the function ofMdTyDcin response to alkaline stress in plants. In our study, it was found that the expression ofMdTyDcwas induced by alkaline stress. Therefore, the apple plants overexpressingMdTyDcwas treated with alkali stress, and we found thatMdTyDcplayed an important role in apple plants’ resistance to alkali stress. Our results showed that the restriction on the growth, the decrease of membrane permeability and the accumulation of Na+were alleviated to various degrees inMdTyDctransgenic plants under alkali stress. In addition, overexpression ofMdTyDcenhanced the root activity and photosynthetic capacity, and improved the enzyme activity related to N metabolism, thus promoting N absorption. It is noteworthy that the dopamine content of these three transgenic lines is significantly higher than that of WT. In summary, these findings indicated thatMdTyDcmay enhance alkaline tolerance of apples by mediating dopamine content, mainly by maintaining high photosynthetic capacity, normal ion homeostasis and strong nitrogen absorption capacity.
- Published
- 2021
29. ImmunoPET imaging of multiple myeloma with [
- Author
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Cheng, Wang, Yumei, Chen, Yun Nan, Hou, Qiufang, Liu, Di, Zhang, Haitao, Zhao, You, Zhang, Shuxian, An, Lianghua, Li, Jian, Hou, Gang, Huang, Jianjun, Liu, Yong Juan, Zhao, and Weijun, Wei
- Subjects
Heterocyclic Compounds, 1-Ring ,Cell Line, Tumor ,Positron-Emission Tomography ,Humans ,Gallium Radioisotopes ,Tissue Distribution ,Multiple Myeloma ,Tomography, X-Ray Computed - Abstract
Multiple myeloma (MM) remains incurable and its diagnosis relies heavily on bone marrow aspiration and biopsy. CD38 is a glycoprotein highly specific for MM. Antibody therapeutics (e.g., daratumumab) targeting CD38 have shown encouraging efficacy in treating MM, either as a monotherapy agent or in combination with other regimens. However, efficient stratification of patients who might benefit from daratumumab therapy and timely monitoring of the therapeutic responses are still clinical challenges. This work aims to devise a CD38-targeted imaging strategy and assess its value in diagnosing MMs.By labeling a CD38-specific single domain antibody (Nb1053) withImmunoPET imaging with [This work successfully developed a novel CD38-targeted immunoPET imaging approach that enabled precise visualization of CD38 and diagnosis of MMs. Upon clinical translation, [
- Published
- 2020
30. The calcium signaling enzyme CD38 - a paradigm for membrane topology defining distinct protein functions
- Author
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Hon Cheung Lee, Qi Wen Deng, and Yong Juan Zhao
- Subjects
Cyclic ADP-Ribose ,Nicotinic acid adenine dinucleotide phosphate ,Physiology ,Chemistry ,Endoplasmic reticulum ,Cell Biology ,ADP-ribosyl Cyclase 1 ,Transmembrane protein ,Cell biology ,chemistry.chemical_compound ,Cytosol ,Membrane topology ,Second messenger system ,Calcium ,Calcium Signaling ,NAD+ kinase ,Lysosomes ,Molecular Biology ,NADP ,Calcium signaling - Abstract
CD38 is a single-pass transmembrane enzyme catalyzing the synthesis of two nucleotide second messengers, cyclic ADP-ribose (cADPR) from NAD and nicotinic acid adenine dinucleotide phosphate (NAADP) from NADP. The former mediates the mobilization of the endoplasmic Ca2+-stores in response to a wide range of stimuli, while NAADP targets the endo-lysosomal stores. CD38 not only possesses multiple enzymatic activities, it also exists in two opposite membrane orientations. Type III CD38 has the catalytic domain facing the cytosol and is responsible for producing cellular cADPR. The type II CD38 has an opposite orientation and is serving as a surface receptor mediating extracellular functions such as cell adhesion and lymphocyte activation. Its ecto-NADase activity also contributes to the recycling of external NAD released by apoptosis. Endocytosis can deliver surface type II CD38 to endo-lysosomes, which acidic environment favors the production of NAADP. This article reviews the rationale and evidence that have led to CD38 as a paradigm for membrane topology defining distinct functions of proteins. Also described is the recent discovery of a hitherto unknown cADPR-synthesizing enzyme, SARM1, ushering in a new frontier in cADPR-mediated Ca2+-signaling.
- Published
- 2022
31. Sepsis-associated Encephalopathy In ICU Admissions: Prevalence, Early Risk of Death, and its Early Prevent and Control
- Author
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Daoming Tong, Yuan-Yuan Gu, Yuan-Wei Wang, Yong-Juan Zhao, Shao-Dan Wang, and Ying Wang
- Subjects
medicine.medical_specialty ,business.industry ,parasitic diseases ,Emergency medicine ,Medicine ,Risk of death ,Sepsis-Associated Encephalopathy ,business - Abstract
Background: Sepsis-associated encephalopathy (SAE) is a common encephalopathy in ICU. We are to definite whether SAE present an high prevalence rate and early risk factors for death in ICU 48 hours, while to cognize its important of early prevention/ control.Methods: We retrospectively enrolled patients with acute critically ill from ICU (January, 2015 to January, 2017). All patients were selected from onset to ICU ≤3 hours. The prevalence and some early risk factors of death in SAE was estimated by using a continuous head and thorax /abdominal cavity CT scans. Results: 748 critically ill adults were analyzed. The prevalence of sepsis within initial 48 hours was 40.4% (302/748). The median time from infection to sepsis was 9 hours ( range, 1-48 ). The SAE (93.4%, 282/302) was diagnosed in sepsis patients, and most of them (96.8%) presented multiple organ dysfunction syndromes (MODS). While the fatality of SAE was in 32.0% at initial 48 hours and 69.1% at initial 14 days. Cox regression analysis, unused antibiotic within initial 3 hours (OR, 0.39; 95% CI, 0.22-0.89), severe inflammatory storm (OR, 0.70; 95% CI, 0.58- 0.94), lower GCS (OR, 2.7; 95% CI, 1.5-3.6), and MODS (OR, 0.37; 95% CI, 0.26-0.96) were early risk factors for death in SAE. Early risk factors for predicting SAE were related to severe inflammatory storm (OR, 3.10; 95% CI, 2.28-4.33), MODS (OR, 3.57; 95% CI, 2.73- 4.67), and unused antibiotics within initial 3 hours (OR, 0.25; 95% CI, 0.11-0.56).Conclusions: SAE in ICU is an high prevalent acute brain dysfunction and most with MODS. The early bad prognosis in SAE was related to the failure of early prevention and control.
- Published
- 2020
32. Structural basis for the inhibition of the papain-like protease of SARS-CoV-2 by small molecules
- Author
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Ziyang Fu, Bin Huang, Jinle Tang, Shuyan Liu, Ming Liu, Yuxin Ye, Zhihong Liu, Yuxian Xiong, Dan Cao, Jihui Li, Xiaogang Niu, Huan Zhou, Yong Juan Zhao, Guoliang Zhang, and Hao Huang
- Subjects
Papain ,chemistry.chemical_compound ,Protease ,Biochemistry ,chemistry ,medicine.medical_treatment ,Virus maturation ,Druggability ,medicine ,ISG15 ,IC50 ,Small molecule ,Cysteine protease - Abstract
SARS-CoV-2 is the pathogen responsible for the COVID-19 pandemic. The SARS-CoV-2 papain-like cysteine protease has been implicated in virus maturation, dysregulation of host inflammation and antiviral immune responses. We showed that PLpro preferably cleaves the K48-ubiquitin linkage while also being capable of cleaving ISG15 modification. The multiple functions of PLpro render it a promising drug target. Therefore, we screened an FDA-approved drug library and also examined available inhibitors against PLpro. Inhibitor GRL0617 showed a promising IC50 of 2.1 μM. The co-crystal structure of SARS-CoV-2 PLpro-C111S in complex with GRL0617 suggests that GRL0617 is a non-covalent inhibitor. NMR data indicate that GRL0617 blocks the binding of ISG15 to PLpro. The antiviral activity of GRL0617 reveal that PLpro is a promising drug target for therapeutically treating COVID-19.One Sentence SummaryCo-crystal structure of PLpro in complex with GRL0617 reveals the druggability of PLpro for SARS-CoV-2 treatment.
- Published
- 2020
33. Resolving the topological enigma in Ca
- Author
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Hon Cheung, Lee and Yong Juan, Zhao
- Subjects
Cyclic ADP-Ribose ,Cytoskeletal Proteins ,JBC Reviews ,Calcium-Binding Proteins ,Animals ,Humans ,Calcium Signaling ,ADP-ribosyl Cyclase 1 ,NADP ,Molecular Chaperones ,RNA, Guide, Kinetoplastida - Abstract
Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) are two structurally distinct messengers that mobilize the endoplasmic and endolysosomal Ca(2+) stores, respectively. Both are synthesized by the CD38 molecule (CD38), which has long been thought to be a type II membrane protein whose catalytic domain, intriguingly, faces to the outside of the cell. Accordingly, for more than 20 years, it has remained unresolved how CD38 can use cytosolic substrates such as NAD and NADP to produce messengers that target intracellular Ca(2+) stores. The discovery of type III CD38, whose catalytic domain faces the cytosol, has now begun to clarify this topological conundrum. This article reviews the ideas and clues leading to the discovery of the type III CD38; highlights an innovative approach for uncovering its natural existence; and discusses the regulators of its activity, folding, and degradation. We also review the compartmentalization of cADPR and NAADP biogenesis. We further discuss the possible mechanisms that promote type III CD38 expression and appraise a proposal of a Ca(2+)-signaling mechanism based on substrate limitation and product translocation. The surprising finding of another enzyme that produces cADPR and NAADP, sterile α and TIR motif–containing 1 (SARM1), is described. SARM1 regulates axonal degeneration and has no sequence similarity with CD38 but can catalyze the same set of multireactions and has the same cytosolic orientation as the type III CD38. The intriguing finding that SARM1 is activated by nicotinamide mononucleotide to produce cADPR and NAADP suggests that it may function as a regulated Ca(2+)-signaling enzyme like CD38.
- Published
- 2019
34. CD38 produces nicotinic acid adenosine dinucleotide phosphate in the lysosome
- Author
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Qi Wen Deng, Ya Jie Chen, Cheng Fang, Ting Li, Yun Nan Hou, Ying Li, Hon Cheung Lee, Yong Juan Zhao, and Guan Jie Xu
- Subjects
0301 basic medicine ,030102 biochemistry & molecular biology ,Chemistry ,media_common.quotation_subject ,hemic and immune systems ,Cell Biology ,Endocytosis ,Biochemistry ,Endolysosome ,Cell biology ,03 medical and health sciences ,Enzyme activator ,030104 developmental biology ,medicine.anatomical_structure ,immune system diseases ,hemic and lymphatic diseases ,Lysosome ,Second messenger system ,medicine ,Internalization ,Molecular Biology ,Intracellular ,media_common ,Calcium signaling - Abstract
Nicotinic acid adenosine dinucleotide phosphate (NAADP) is a Ca2+-mobilizing second messenger that regulates a wide range of biological activities. However, the mechanism of its biogenesis remains controversial. CD38 is the only enzyme known to catalyze NAADP synthesis from NADP and nicotinic acid. CD38-mediated catalysis requires an acidic pH, suggesting that NAADP may be produced in acidic endolysosomes, but this hypothesis is untested. In this study, using human cell lines, we specifically directed CD38 to the endolysosomal system and assessed cellular NAADP production. First, we found that nanobodies targeting various epitopes on the C-terminal domain of CD38 could bind to cell surface–localized CD38 and induce its endocytosis. We also found that CD38 internalization occurred via a clathrin-dependent pathway, delivered CD38 to the endolysosome, and elevated intracellular NAADP levels. We also created a CD38 variant for lysosome-specific expression, which not only withstood the degradative environment in the lysosome, but was also much more active than WT CD38 in elevating cellular NAADP levels. Supplementing CD38-expressing cells with nicotinic acid substantially increased cellular NAADP levels. These results demonstrate that endolysosomal CD38 can produce NAADP in human cells. They further suggest that CD38's compartmentalization to the lysosome may allow for its regulation via substrate access, rather than enzyme activation, thereby providing a reliable mechanism for regulating cellular NAADP production.
- Published
- 2018
35. Development of Stabilized Peptide-Based PROTACs against Estrogen Receptor α
- Author
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Qiwen Deng, Zigang Li, Yanhong Jiang, Mingsheng Xie, Feng Yin, Weihao Zheng, Yong Juan Zhao, Longjian Chen, Xuan Qin, and Hui Zhao
- Subjects
0301 basic medicine ,Transcription, Genetic ,Ubiquitin-Protein Ligases ,Estrogen receptor ,Breast Neoplasms ,Peptide ,Cell-Penetrating Peptides ,01 natural sciences ,Biochemistry ,Pentapeptide repeat ,Mice ,03 medical and health sciences ,Chimera (genetics) ,Transcription (biology) ,Cell Line, Tumor ,Aspartic acid ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Cell Proliferation ,chemistry.chemical_classification ,biology ,010405 organic chemistry ,Chemistry ,Estrogen Receptor alpha ,General Medicine ,0104 chemical sciences ,Ubiquitin ligase ,Cell biology ,Cross-Linking Reagents ,030104 developmental biology ,Cell culture ,Drug Design ,MCF-7 Cells ,biology.protein ,Heterografts ,Molecular Medicine ,Female - Abstract
Peptide modulators targeting protein-protein interactions (PPIs) exhibit greater potential than small-molecule drugs in several important aspects including facile modification and relative large contact surface area. Stabilized peptides constructed by variable chemistry methods exhibit improved peptide stability and cell permeability compared to that of the linears. Herein, we designed a stabilized peptide-based proteolysis-targeting chimera (PROTAC) targeting estrogen receptor α (ERα) by tethering an N-terminal aspartic acid cross-linked stabilized peptide ERα modulator (TD-PERM) with a pentapeptide that binds the Von Hippel-Lindau (VHL) E3 ubiquitin ligase complex. The resulting heterobifunctional peptide (TD-PROTAC) selectively recruits ERα to the VHL E3 ligase complex, leading to the degradation of ERα in a proteasome-dependent manner. Compared with the control peptides, TD-PROTAC shows significantly enhanced activities in reducing the transcription of the ERα-downstream genes and inhibiting the proliferation of ERα-positive breast cancer cells. In addition, in vivo experiments indicate that TD-PROTAC leads to tumor regression in the MCF-7 mouse xenograft model. This work is a successful attempt to construct PROTACs based on cell-permeable stabilized peptides, which significantly broadens the chemical space of PROTACs and stabilized peptides.
- Published
- 2017
36. Quantitative proteomics reveals arsenic attenuates stem‐loop binding protein stability via a chaperone complex containing heat shock proteins and ERp44
- Author
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Jingzi Zhang, Yan Guo, Chunyuan Jin, Bing-Hao Wang, Wei Ming He, Yibing Ding, Chao-Jun Li, Yong Juan Zhao, Ruoyu Zhao, Yue Zhao, Lei Fang, Wei Ren, and Danqi Chen
- Subjects
Proteomics ,chemistry.chemical_element ,Biochemistry ,Arsenic ,03 medical and health sciences ,Heat shock protein ,Humans ,Molecular Biology ,Heat-Shock Proteins ,030304 developmental biology ,mRNA Cleavage and Polyadenylation Factors ,SLBP ,0303 health sciences ,Gene knockdown ,integumentary system ,biology ,Protein Stability ,Chemistry ,Binding protein ,030302 biochemistry & molecular biology ,Membrane Proteins ,Nuclear Proteins ,Cell cycle ,Cell biology ,Histone ,biology.protein ,Chaperone complex ,Molecular Chaperones ,Protein Binding - Abstract
Arsenic pollution impacts health of millions of people in the world. Inorganic arsenic is a carcinogenic agent in skin and lung cancers. The stem-loop binding protein (SLBP) binds to the stem-loop of the canonical histone mRNA and regulates its metabolism during cell cycle. Our previous work has shown arsenic induces ubiquitin-proteasome dependent degradation of SLBP and contributes to lung cancer. In this study, we established the first comprehensive SLBP interaction network by affinity purification-mass spectrometry (AP-MS) analysis, and further demonstrated arsenic enhanced the association between SLBP and a crucial chaperone complex containing heat shock proteins (HSPs) and ERp44. Strikingly, knockdown of these proteins markedly rescued the protein level of SLBP under arsenic exposure conditions, and abolished the increasing migration capacity of BEAS-2B cells induced by arsenic. Taken together, our study provides a potential new mechanism that a chaperone complex containing HSPs and ERp44 attenuates the stability of SLBP under both normal and arsenic exposure conditions, which could be essential for arsenic-induced high cell migration. This article is protected by copyright. All rights reserved.
- Published
- 2021
37. Cytosolic interaction of type III human CD38 with CIB1 modulates cellular cyclic ADP-ribose levels
- Author
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Song Lu Li, Cheng Fang, Wan Hua Li, Hon Cheung Lee, Yong Juan Zhao, Zhi Ying Zhao, Yun Nan Hou, Jun Liu, and Ting Li
- Subjects
0301 basic medicine ,Multidisciplinary ,Immunoprecipitation ,Endoplasmic reticulum ,Biological Sciences ,Biology ,CD38 ,Cyclic ADP-ribose ,Cell biology ,Small hairpin RNA ,03 medical and health sciences ,Cytosol ,Bimolecular fluorescence complementation ,chemistry.chemical_compound ,030104 developmental biology ,chemistry ,Biochemistry ,Calcium signaling - Abstract
CD38 catalyzes the synthesis of the Ca2+ messenger, cyclic ADP-ribose (cADPR). It is generally considered to be a type II protein with the catalytic domain facing outside. How it can catalyze the synthesis of intracellular cADPR that targets the endoplasmic Ca2+ stores has not been resolved. We have proposed that CD38 can also exist in an opposite type III orientation with its catalytic domain facing the cytosol. Here, we developed a method using specific nanobodies to immunotarget two different epitopes simultaneously on the catalytic domain of the type III CD38 and firmly established that it is naturally occurring in human multiple myeloma cells. Because type III CD38 is topologically amenable to cytosolic regulation, we used yeast-two-hybrid screening to identify cytosolic Ca2+ and integrin-binding protein 1 (CIB1), as its interacting partner. The results from immunoprecipitation, ELISA, and bimolecular fluorescence complementation confirmed that CIB1 binds specifically to the catalytic domain of CD38, in vivo and in vitro. Mutational studies established that the N terminus of CIB1 is the interacting domain. Using shRNA to knock down and Cas9/guide RNA to knock out CIB1, a direct correlation between the cellular cADPR and CIB1 levels was demonstrated. The results indicate that the type III CD38 is functionally active in producing cellular cADPR and that the activity is specifically modulated through interaction with cytosolic CIB1.
- Published
- 2017
38. Luteolinidin Protects the Postischemic Heart through CD38 Inhibition with Preservation of NAD(P)(H)
- Author
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Craig Hemann, Hon Cheung Lee, James Boslett, Yong Juan Zhao, and Jay L. Zweier
- Subjects
Male ,0301 basic medicine ,Cardiotonic Agents ,Endothelium ,Myocardial Ischemia ,Vasodilation ,Pharmacology ,Cardiovascular ,Nitric oxide ,Anthocyanins ,Rats, Sprague-Dawley ,Luteolinidin ,03 medical and health sciences ,chemistry.chemical_compound ,Enos ,medicine ,Animals ,Humans ,Endothelial dysfunction ,Membrane Glycoproteins ,Dose-Response Relationship, Drug ,biology ,Tetrahydrobiopterin ,medicine.disease ,biology.organism_classification ,ADP-ribosyl Cyclase 1 ,Recombinant Proteins ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Biochemistry ,Molecular Medicine ,NAD+ kinase ,NADP ,medicine.drug - Abstract
We recently showed that ischemia/reperfusion (I/R) of the heart causes CD38 activation with resultant depletion of the cardiac NADP(H) pool, which is most marked in the endothelium. This NADP(H) depletion was shown to limit the production of nitric oxide by endothelial nitric oxide synthase (eNOS), which requires NADPH for nitric oxide production, resulting in greatly altered endothelial function. Therefore, intervention with CD38 inhibitors could reverse postischemic eNOS-mediated endothelial dysfunction. Here, we evaluated the potency of the CD38 inhibitor luteolinidin, an anthocyanidin, at blocking CD38 activity and preserving endothelial and myocardial function in the postischemic heart. Initially, we characterized luteolinidin as a CD38 inhibitor in vitro to determine its potency and mechanism of inhibition. We then tested luteolinidin in the ex vivo isolated heart model, where we determined luteolinidin uptake with aqueous and liposomal delivery methods. Optimal delivery methods were then further tested to determine the effect of luteolinidin on postischemic NAD(P)(H) and tetrahydrobiopterin levels. Finally, through nitric oxide synthase–dependent coronary flow and left ventricular functional measurements, we evaluated the efficacy of luteolinidin to protect vascular and contractile function, respectively, after I/R. With enhanced postischemic preservation of NADPH and tetrahydrobiopterin, there was a dose-dependent effect of luteolinidin on increasing recovery of endothelium-dependent vasodilatory function, as well as enhancing the recovery of left ventricular contractile function with increased myocardial salvage. Thus, luteolinidin is a potent CD38 inhibitor that protects the heart against I/R injury with preservation of eNOS function and prevention of endothelial dysfunction.
- Published
- 2017
39. A Cell-Permeant Mimetic of NMN Activates SARM1 to Produce Cyclic ADP-Ribose and Induce Non-apoptotic Cell Death
- Author
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Zhengshuang Xu, Lihe Zhang, Ben Zhang, Zhe Chen, Yong Juan Zhao, Xu Jie Xie, Liangren Zhang, Jun Liu, Song Lu Li, Ting Li, Jun Gang Lu, Zhi Ying Zhao, Hon Cheung Lee, and Wan Hua Li
- Subjects
0301 basic medicine ,Conformational change ,Cell ,Endogeny ,02 engineering and technology ,CD38 ,Cyclic ADP-ribose ,Biochemistry ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,medicine ,lcsh:Science ,chemistry.chemical_classification ,Multidisciplinary ,Activator (genetics) ,Chemistry ,Biochemical Mechanism ,021001 nanoscience & nanotechnology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Enzyme ,Enzymology ,lcsh:Q ,NAD+ kinase ,0210 nano-technology - Abstract
Summary SARM1, an NAD-utilizing enzyme, regulates axonal degeneration. We show that CZ-48, a cell-permeant mimetic of NMN, activated SARM1 in vitro and in cellulo to cyclize NAD and produce a Ca2+ messenger, cADPR, with similar efficiency as NMN. Knockout of NMN-adenylyltransferase elevated cellular NMN and activated SARM1 to produce cADPR, confirming NMN was its endogenous activator. Determinants for the activating effects and cell permeability of CZ-48 were identified. CZ-48 activated SARM1 via a conformational change of the auto-inhibitory domain and dimerization of its catalytic domain. SARM1 catalysis was similar to CD38, despite having no sequence similarity. Both catalyzed similar set of reactions, but SARM1 had much higher NAD-cyclizing activity, making it more efficient in elevating cADPR. CZ-48 acted selectively, activating SARM1 but inhibiting CD38. In SARM1-overexpressing cells, CZ-48 elevated cADPR, depleted NAD and ATP, and induced non-apoptotic death. CZ-48 is a specific modulator of SARM1 functions in cells., Graphical Abstract, Highlights • CZ-48, a cell-permeant mimetic of NMN, activates SARM1 but inhibits CD38 enzymatically • SARM1 catalysis is similar to CD38, but with higher cyclase activity • Activation by CZ-48 or NMN elicits conformational changes in SARM1 • Activation of SARM1 causes cADPR production, NAD depletion, and non-apoptotic cell death, Biochemistry; Enzymology; Biochemical Mechanism
- Published
- 2019
40. Solution-phase total synthesis of teixobactin
- Author
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Zhengshuang Xu, Yun Nan Hou, Sigui Chen, Tao Ye, Bowen Gao, and Yong Juan Zhao
- Subjects
chemistry.chemical_classification ,010405 organic chemistry ,Chemistry ,Longest linear sequence ,Stereochemistry ,Organic Chemistry ,Teixobactin ,Total synthesis ,Peptide ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Coupling reaction ,0104 chemical sciences ,chemistry.chemical_compound ,Yield (chemistry) ,Stereoselectivity ,Physical and Theoretical Chemistry ,Derivative (chemistry) - Abstract
The first solution-phase total synthesis of the cyclic depsipeptide teixobactin is described. Stereoselective construction of l-allo-enduracididine was established, and the protective groups for the peptide coupling reactions and conditions for the assembly of the fragments were also optimised. The longest linear sequence for the total synthesis was 20 steps from the known l-cis-4-hydroxyproline derivative and gave a 5.6% overall yield. This solution-phase total synthesis could serve as a complement to the current solid-phase synthesis of teixobactin.
- Published
- 2019
41. Direct Gating of the TRPM2 Channel by cADPR via Specific Interactions with the ADPR Binding Pocket
- Author
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Huan Liu, Lin-Hua Jiang, Peiwu Ye, Fan Yang, Liangren Zhang, Xiwen Xue, Lixin Yang, Yang Wu, Peilin Yu, Yong Juan Zhao, Zhongtang Li, Zhenming Liu, Jianhong Luo, Wei Yang, Cheng Fang, Lihe Zhang, and Xiafei Yu
- Subjects
0301 basic medicine ,Protein Conformation ,Binding pocket ,TRPM Cation Channels ,Gating ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Humans ,Point Mutation ,TRPM2 ,Surface plasmon resonance ,Pyrophosphatases ,ADP-ribosyl Cyclase ,lcsh:QH301-705.5 ,Adenosine Diphosphate Ribose ,Binding Sites ,Activator (genetics) ,Chemistry ,Electrophysiology ,030104 developmental biology ,HEK293 Cells ,lcsh:Biology (General) ,Biophysics ,Calcium ,030217 neurology & neurosurgery - Abstract
Summary: cADPR is a well-recognized signaling molecule by modulating the RyRs, but considerable debate exists regarding whether cADPR can bind to and gate the TRPM2 channel, which mediates oxidative stress signaling in diverse physiological and pathological processes. Here, we show that purified cADPR evoked TRPM2 channel currents in both whole-cell and cell-free single-channel recordings and specific binding of cADPR to the purified NUDT9-H domain of TRPM2 by surface plasmon resonance. Furthermore, by combining computational modeling with electrophysiological recordings, we show that the TRPM2 channels carrying point mutations at H1346, T1347, L1379, S1391, E1409, and L1484 possess distinct sensitivity profiles for ADPR and cADPR. These results clearly indicate cADPR is a bona fide activator at the TRPM2 channel and clearly delineate the structural basis for cADPR binding, which not only lead to a better understanding in the gating mechanism of TRPM2 channel but also shed light on a cADPR-induced RyRs-independent Ca2+ signaling mechanism. : Yu et al. demonstrate that cADPR, which was thought to be an intracellular calcium messenger by modulating function of the ryanodine receptors (RyRs), also activates the TRPM2 channel. Their results suggest that cADPR regulates intracellular calcium release through multiple pathways. Keywords: cADPR, NUDT9-H domain, TRPM2 channel
- Published
- 2018
42. Permeant fluorescent probes visualize the activation of SARM1 and uncover an antineurodegenerative drug candidate.
- Author
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Wan Hua Li, Ke Huang, Yang Cai, Qian Wen Wang, Wen Jie Zhu, Yun Nan Hou, Sujing Wang, Sheng Cao, Zhi Ying Zhao, Xu Jie Xie, Yang Du, Chi-Sing Lee, Hon Cheung Lee, Hongmin Zhang, and Yong Juan Zhao
- Published
- 2021
- Full Text
- View/download PDF
43. Correction: CD38 produces nicotinic acid adenine dinucleotide phosphate in the lysosome
- Author
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Yun Nan Hou, Qi Wen Deng, Yong Juan Zhao, Cheng Fang, Ying Li, Hon Cheung Lee, Ting Li, Ya Jie Chen, and Guan Jie Xu
- Subjects
Vasodilator Agents ,CD38 ,Niacin ,Biochemistry ,chemistry.chemical_compound ,immune system diseases ,hemic and lymphatic diseases ,Lysosome ,medicine ,Humans ,Calcium Signaling ,Molecular Biology ,Membrane Glycoproteins ,Nicotinic acid adenine dinucleotide phosphate ,hemic and immune systems ,Cell Biology ,Single-Domain Antibodies ,ADP-ribosyl Cyclase 1 ,Endocytosis ,HEK293 Cells ,medicine.anatomical_structure ,chemistry ,Additions and Corrections ,Calcium ,Lysosomes ,NADP ,HeLa Cells - Abstract
Nicotinic acid adenosine dinucleotide phosphate (NAADP) is a Ca(2+)-mobilizing second messenger that regulates a wide range of biological activities. However, the mechanism of its biogenesis remains controversial. CD38 is the only enzyme known to catalyze NAADP synthesis from NADP and nicotinic acid. CD38-mediated catalysis requires an acidic pH, suggesting that NAADP may be produced in acidic endolysosomes, but this hypothesis is untested. In this study, using human cell lines, we specifically directed CD38 to the endolysosomal system and assessed cellular NAADP production. First, we found that nanobodies targeting various epitopes on the C-terminal domain of CD38 could bind to cell surface–localized CD38 and induce its endocytosis. We also found that CD38 internalization occurred via a clathrin-dependent pathway, delivered CD38 to the endolysosome, and elevated intracellular NAADP levels. We also created a CD38 variant for lysosome-specific expression, which not only withstood the degradative environment in the lysosome, but was also much more active than WT CD38 in elevating cellular NAADP levels. Supplementing CD38-expressing cells with nicotinic acid substantially increased cellular NAADP levels. These results demonstrate that endolysosomal CD38 can produce NAADP in human cells. They further suggest that CD38's compartmentalization to the lysosome may allow for its regulation via substrate access, rather than enzyme activation, thereby providing a reliable mechanism for regulating cellular NAADP production.
- Published
- 2019
44. [Changes of ion absorption, distribution and essential oil components of flowering Schizonepeta tenuifolia under salt stress]
- Author
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Ying, Zhou, Li-Jin, Huang, Yong-Juan, Zhao, Nan-Yu, Tang, Ren-Jun, Qu, Xiao-Qing, Tang, and Kang-Cai, Wang
- Subjects
Plant Leaves ,Lamiaceae ,Sodium ,Oils, Volatile ,Sodium Chloride ,Salt Stress - Abstract
In this paper, a pot experiment using quartz sands was conducted to study the effects of different concentrations of NaCl (0, 25, 50, 75, 100 mmol·L⁻¹) on the ion absorption, distribution and essential oil components of flowering Schizonepeta tenuifolia. The results showed that as NaCl concentration increased, Na⁺ content of root, stem, leaf and flower increased significantly, and that of the aerial parts was in a higher level than in the root. Regarding the K⁺ content, it decreased in the root but increased in stem, leaf and flower. Some changes were detected in the Ca²⁺ content, but not significant on the whole. The value of K⁺/Na⁺ and Ca²⁺/Na⁺ reduced as a result of increasing NaCl concentrations. The content of essential oil increased under medium salt treatment (50 mmol·L⁻¹ NaCl). However, the synthesis and accumulation of essential oil was inhibited by the serious salt treatment (100 mmol·L⁻¹ NaCl). Over 98% of the essential oil components were terpenes, in which pulegone and menthone were the most two abundant compounds. Varieties of essential oil components did not change significantly under salt stress but their relative proportions did. The transformation of pulegone to menthone was enhanced and the value of pulegone/menthone based on their relative contents decreased with NaCl concentration increasing. Consequently, menthone ranked the most abundant compound by replacing pulegone. Relative content of D-limonene increased under medium and serious salt stress, and that of β-caryophyllene only increased under mild treatments. So our research could provide references for the standard cultivation on saline soil of S. tenuifolia.
- Published
- 2018
45. Nanobody-based dual epitopes protein identification (DepID) assay for measuring soluble CD38 in plasma of multiple myeloma patients
- Author
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Cheng Fang, Hon Cheung Lee, Song Lu Li, Yun Nan Hou, Qiaoxia Zhang, Xin Du, Yong Juan Zhao, and Ting Li
- Subjects
0301 basic medicine ,Models, Molecular ,Protein Conformation ,CD38 ,Biochemistry ,Epitope ,Analytical Chemistry ,03 medical and health sciences ,Epitopes ,Antigen ,Limit of Detection ,Cell Line, Tumor ,medicine ,Environmental Chemistry ,Potency ,Humans ,Luciferase ,Spectroscopy ,Multiple myeloma ,Chemistry ,Plasma levels ,Single-Domain Antibodies ,medicine.disease ,Molecular biology ,ADP-ribosyl Cyclase 1 ,030104 developmental biology ,Solubility ,Disease Progression ,Protein identification ,Multiple Myeloma ,Blood Chemical Analysis - Abstract
Background CD38 is a surface membrane antigen highly expressed in malignant blood cells, such as multiple myeloma (MM). A soluble form of CD38 (sCD38) is also present in the plasma, deriving likely from the shedding from the cells. The plasma levels of sCD38 should thus correlate closely with the proliferation of the MM cells, allowing the development of a simple diagnostic blood test for monitoring the progress of the disease. However, the plasma sCD38 levels are extremely low, requiring the design of a highly sensitive and specific assay. Results In this study, we developed an ultra-sensitive assay, based on two nanobodies (Nbs) targeting two distinct epitopes of sCD38. One Nb acts as a capturer, and the other is fused with the firefly luciferase serving as a reporter to ensure sensitivity. We showed that this D ual e pitopes p rotein ID entification (DepID) assay has sensitivity reaching 10 pg/mL, which is 10 times higher than that of a commercial ELISA kit. By this method, we were able to precisely quantify the levels of sCD38 in the plasma of MM patients, which were significantly higher than those from healthy donors. We further showed that the increase plasma levels of sCD38 correlated with the progress of MM. Conclusion We have developed a Nb-based luminescence sandwich assay, named as DepID, for quantification of the soluble CD38 in MM patients' plasma and showed the potency of this method as a tool for general diagnosis of MM or companion diagnosis of the CD38-targeted therapies.
- Published
- 2018
46. Design, Synthesis and SAR Studies of NAD Analogues as Potent Inhibitors towards CD38 NADase
- Author
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Jianguo Li, Hon Cheung Lee, Yong Juan Zhao, Kehui Zhang, Shengjun Wang, Xuan Wang, Wenjie Zhu, Liangren Zhang, and Lihe Zhang
- Subjects
synthesis ,Stereochemistry ,Guanine ,Pharmaceutical Science ,Chemistry Techniques, Synthetic ,CD38 ,Nicotinamide adenine dinucleotide ,Article ,Cofactor ,Substrate Specificity ,Analytical Chemistry ,lcsh:QD241-441 ,chemistry.chemical_compound ,lcsh:Organic chemistry ,inhibitors ,Drug Discovery ,Enzyme Inhibitors ,Physical and Theoretical Chemistry ,chemistry.chemical_classification ,Molecular Structure ,biology ,Organic Chemistry ,NAD ,NAD analogues ,ADP-ribosyl Cyclase 1 ,Guanine Nucleotides ,Enzyme Activation ,Enzyme ,Glycerol-3-phosphate dehydrogenase ,Biochemistry ,chemistry ,Chemistry (miscellaneous) ,Drug Design ,Second messenger system ,biology.protein ,Molecular Medicine ,NAD+ kinase ,Protein Binding - Abstract
Nicotinamide adenine dinucleotide (NAD), one of the most important coenzymes in the cells, is a substrate of the signaling enzyme CD38, by which NAD is converted to a second messenger, cyclic ADP-ribose, which releases calcium from intracellular calcium stores. Starting with 2′-deoxy-2′-fluoroarabinosyl-β-nicotinamide adenine dinucleotide (ara-F NAD), a series of NAD analogues were synthesized and their activities to inhibit CD38 NAD glycohydrolase (NADase) were evaluated. The adenosine-modified analogues showed potent inhibitory activities, among which 2′-deoxy-2′-fluoroarabinosyl-β-nicotinamide guanine dinucleotide (ara-F NGD) was the most effective one. The structure-activity relationship of NAD analogues was also discussed.
- Published
- 2014
47. Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38
- Author
-
William, Fumey, Julia, Koenigsdorf, Valentin, Kunick, Stephan, Menzel, Kerstin, Schütze, Mandy, Unger, Levin, Schriewer, Friedrich, Haag, Gerhard, Adam, Anna, Oberle, Mascha, Binder, Ralf, Fliegert, Andreas, Guse, Yong Juan, Zhao, Hon, Cheung Lee, Fabio, Malavasi, Fernando, Goldbaum, Rob, van Hegelsom, Catelijne, Stortelers, Peter, Bannas, and Friedrich, Koch-Nolte
- Subjects
Membrane Glycoproteins ,Antibodies, Monoclonal ,Mice, Nude ,Antineoplastic Agents ,Single-Domain Antibodies ,ADP-ribosyl Cyclase 1 ,Xenograft Model Antitumor Assays ,Article ,Disease Models, Animal ,Epitopes ,Mice ,immune system diseases ,hemic and lymphatic diseases ,Cell Line, Tumor ,Animals ,Humans ,Cell Surface Display Techniques ,Multiple Myeloma ,Camelids, New World ,Fluorescent Dyes - Abstract
The cell surface ecto-enzyme CD38 is a promising target antigen for the treatment of hematological malignancies, as illustrated by the recent approval of daratumumab for the treatment of multiple myeloma. Our aim was to evaluate the potential of CD38-specific nanobodies as novel diagnostics for hematological malignancies. We successfully identified 22 CD38-specific nanobody families using phage display technology from immunized llamas. Crossblockade analyses and in-tandem epitope binning revealed that the nanobodies recognize three different non-overlapping epitopes, with four nanobody families binding complementary to daratumumab. Three nanobody families inhibit the enzymatic activity of CD38 in vitro, while two others were found to act as enhancers. In vivo, fluorochrome-conjugated CD38 nanobodies efficiently reach CD38 expressing tumors in a rodent model within 2 hours after intravenous injection, thereby allowing for convenient same day in vivo tumor imaging. These nanobodies represent highly specific tools for modulating the enzymatic activity of CD38 and for diagnostic monitoring CD38-expressing tumors.
- Published
- 2017
48. Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo
- Author
-
Friedrich Koch-Nolte, Ralf Fliegert, Kerstin Schütze, Mandy Unger, Friedrich Haag, Valentin Kunick, Fernando Alberto Goldbaum, Andreas H. Guse, Fabio Malavasi, William Fumey, Peter Bannas, Hon Cheung Lee, Rob van Hegelsom, Levin Schriewer, Yong Juan Zhao, Stephan Menzel, Anna Oberle, Gerhard Adam, Julia Koenigsdorf, Mascha Binder, and Catelijne Stortelers
- Subjects
0301 basic medicine ,Phage display ,CIENCIAS MÉDICAS Y DE LA SALUD ,Cell ,lcsh:Medicine ,Biology ,Neutralizing antibodies ,Epitope ,Imaging ,Biotecnología de la Salud ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,immune system diseases ,hemic and lymphatic diseases ,Epitope binning ,medicine ,purl.org/becyt/ford/3.4 [https] ,lcsh:Science ,Multidisciplinary ,lcsh:R ,Daratumumab ,In vitro ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Nanobodies ,purl.org/becyt/ford/3 [https] ,lcsh:Q ,Antibody ,CD38 ,Otras Biotecnologías de la Salud - Abstract
The cell surface ecto-enzyme CD38 is a promising target antigen for the treatment of hematological malignancies, as illustrated by the recent approval of daratumumab for the treatment of multiple myeloma. Our aim was to evaluate the potential of CD38-specific nanobodies as novel diagnostics for hematological malignancies. We successfully identified 22 CD38-specific nanobody families using phage display technology from immunized llamas. Crossblockade analyses and in-tandem epitope binning revealed that the nanobodies recognize three different non-overlapping epitopes, with four nanobody families binding complementary to daratumumab. Three nanobody families inhibit the enzymatic activity of CD38 in vitro, while two others were found to act as enhancers. In vivo, fluorochrome-conjugated CD38 nanobodies efficiently reach CD38 expressing tumors in a rodent model within 2 hours after intravenous injection, thereby allowing for convenient same day in vivo tumor imaging. These nanobodies represent highly specific tools for modulating the enzymatic activity of CD38 and for diagnostic monitoring CD38-expressing tumors. Fil: Fumey, William. University Medical Center Hamburg-Eppendorf; Alemania Fil: Koenigsdorf, Julia. University Medical Center Hamburg-Eppendorf; Alemania Fil: Kunick, Valentin. University Medical Center Hamburg-Eppendorf; Alemania Fil: Menzel, Stephan. University Medical Center Hamburg-Eppendorf; Alemania Fil: Schütze, Kerstin. University Medical Center Hamburg-Eppendorf; Alemania Fil: Unger, Mandy. University Medical Center Hamburg-Eppendorf; Alemania Fil: Schriewer, Levin. University Medical Center Hamburg-Eppendorf; Alemania Fil: Haag, Friedrich. University Medical Center Hamburg-Eppendorf; Alemania Fil: Adam, Gerhard. University Medical Center Hamburg-Eppendorf; Alemania Fil: Oberle, Anna. University Medical Center Hamburg-Eppendorf; Alemania Fil: Binder, Mascha. University Medical Center Hamburg-Eppendorf; Alemania Fil: Fliegert, Ralf. University Medical Center Hamburg-Eppendorf; Alemania Fil: Guse, Andreas. University Medical Center Hamburg-Eppendorf; Alemania Fil: Zhao, Yong Juan. Peking University; China Fil: Lee, Hon Cheung. Peking University; China Fil: Malavasi, Fabio. Università di Torino; Italia Fil: Goldbaum, Fernando Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación Instituto Leloir; Argentina Fil: Van Hegelsom, Rob. Ablynx nv; Bélgica Fil: Stortelers, Catelijne. Ablynx nv; Bélgica Fil: Bannas, Peter. University Medical Center Hamburg-Eppendorf; Alemania Fil: Koch Nolte, Friedrich. University Medical Center Hamburg-Eppendorf; Alemania
- Published
- 2017
49. The Type III Calcium Signaling Mechanism of CD38
- Author
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Yong Juan Zhao and Hon Cheung Lee
- Subjects
Chemistry ,Materials Science (miscellaneous) ,CD38 ,Mechanism (sociology) ,Calcium signaling ,Cell biology - Published
- 2014
50. Resolving the topological enigma in Ca2+ signaling by cyclic ADP-ribose and NAADP.
- Author
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Hon Cheung Lee and Yong Juan Zhao
- Subjects
- *
NIACIN , *CD38 antigen , *CATALYTIC domains , *MEMBRANE proteins , *ADENOSINE diphosphate , *RIDDLES , *NICOTINAMIDE adenine dinucleotide phosphate - Abstract
Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) are two structurally distinct messengers that mobilize the endoplasmic and endolysosomal Ca2+ stores, respectively. Both are synthesized by the CD38 molecule (CD38), which has long been thought to be a type II membrane protein whose catalytic domain, intriguingly, faces to the outside of the cell. Accordingly, for more than 20 years, it has remained unresolved how CD38 can use cytosolic substrates such as NAD and NADP to produce messengers that target intracellular Ca2+ stores. The discovery of type III CD38, whose catalytic domain faces the cytosol, has now begun to clarify this topological conundrum. This article reviews the ideas and clues leading to the discovery of the type III CD38; highlights an innovative approach for uncovering its natural existence; and discusses the regulators of its activity, folding, and degradation. We also review the compartmentalization of cADPR and NAADP biogenesis. We further discuss the possible mechanisms that promote type III CD38 expression and appraise a proposal of a Ca2+-signaling mechanism based on substrate limitation and product translocation. The surprising finding of another enzyme that produces cADPR and NAADP, sterile α and TIR motif-containing 1 (SARM1), is described. SARM1 regulates axonal degeneration and has no sequence similarity with CD38 but can catalyze the same set of multireactions and has the same cytosolic orientation as the type III CD38. The intriguing finding thatSARM1is activated by nicotinamide mononucleotide to produce cADPR and NAADP suggests that it may function as a regulated Ca2+-signaling enzyme like CD38. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
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