254 results on '"Hon Cheung Lee"'
Search Results
2. Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative 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
- Subjects
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
- Full Text
- View/download PDF
3. 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
- Subjects
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
- Published
- 2019
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4. Design, Synthesis and SAR Studies of NAD Analogues as Potent Inhibitors towards CD38 NADase
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Shengjun Wang, Wenjie Zhu, Xuan Wang, Jianguo Li, Kehui Zhang, Liangren Zhang, Yong-Juan Zhao, Hon Cheung Lee, and Lihe Zhang
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synthesis ,NAD analogues ,CD38 ,inhibitors ,Organic chemistry ,QD241-441 - 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
- Full Text
- View/download PDF
5. 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
- Subjects
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.
- Published
- 2016
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- View/download PDF
6. CD38 Structure-Based Inhibitor Design Using the N1-Cyclic Inosine 5'-Diphosphate Ribose Template.
- Author
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Christelle Moreau, Qun Liu, Richard Graeff, Gerd K Wagner, Mark P Thomas, Joanna M Swarbrick, Satoshi Shuto, Hon Cheung Lee, Quan Hao, and Barry V L Potter
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Medicine ,Science - Abstract
Few inhibitors exist for CD38, a multifunctional enzyme catalyzing the formation and metabolism of the Ca(2+)-mobilizing second messenger cyclic adenosine 5'-diphosphoribose (cADPR). Synthetic, non-hydrolyzable ligands can facilitate structure-based inhibitor design. Molecular docking was used to reproduce the crystallographic binding mode of cyclic inosine 5'-diphosphoribose (N1-cIDPR) with CD38, revealing an exploitable pocket and predicting the potential to introduce an extra hydrogen bond interaction with Asp-155. The purine C-8 position of N1-cIDPR (IC50 276 µM) was extended with an amino or diaminobutane group and the 8-modified compounds were evaluated against CD38-catalyzed cADPR hydrolysis. Crystallography of an 8-amino N1-cIDPR:CD38 complex confirmed the predicted interaction with Asp-155, together with a second H-bond from a realigned Glu-146, rationalizing the improved inhibition (IC50 56 µM). Crystallography of a complex of cyclic ADP-carbocyclic ribose (cADPcR, IC50 129 µM) with CD38 illustrated that Glu-146 hydrogen bonds with the ligand N6-amino group. Both 8-amino N1-cIDPR and cADPcR bind deep in the active site reaching the catalytic residue Glu-226, and mimicking the likely location of cADPR during catalysis. Substantial overlap of the N1-cIDPR "northern" ribose monophosphate and the cADPcR carbocyclic ribose monophosphate regions suggests that this area is crucial for inhibitor design, leading to a new compound series of N1-inosine 5'-monophosphates (N1-IMPs). These small fragments inhibit hydrolysis of cADPR more efficiently than the parent cyclic compounds, with the best in the series demonstrating potent inhibition (IC50 = 7.6 µM). The lower molecular weight and relative simplicity of these compounds compared to cADPR make them attractive as a starting point for further inhibitor design.
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- 2013
- Full Text
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7. 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
8. ENZYMATIC AND ENDOGENOUS SYNTHESIS OF NAD(P)‐DERIVED CALCIUM‐MOBILIZING MESSENGERS
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Hon Cheung Lee and Yong Juan Zhao
- Published
- 2022
9. GALA peptide improves the potency of nanobody–drug conjugates by lipid-induced helix formation
- Author
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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.
- Published
- 2021
10. Resolving the topological enigma in Ca2+ signaling by cyclic ADP-ribose and NAADP
- Author
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Yong Juan Zhao and Hon Cheung Lee
- Subjects
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.
- Published
- 2019
11. The transferrin receptor CD71 regulates type II CD38, revealing tight topological compartmentalization of intracellular cyclic ADP-ribose production
- Author
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Lei Fang, Qi Wen Deng, Hon Cheung Lee, Wei Ming He, Ting Li, Yong Juan Zhao, and Jingzi Zhang
- Subjects
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.
- Published
- 2019
12. Author response: Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate
- Author
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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
13. Rational Design and Identification of Small‐Molecule Allosteric Inhibitors of CD38
- Author
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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
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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
14. 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
15. Anti-Multiple Myeloma Activity of Nanobody-Based Anti-CD38 Chimeric Antigen Receptor T Cells
- Author
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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
- Subjects
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
16. 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
17. 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
18. 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
19. 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
20. 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
21. 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
22. 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
23. Fluorescent analogs of cyclic ADP-ribose: synthesis, spectral characterization, and use
- Author
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Graeff, Richard M., Walseth, Timothy F., Hill, Heather K., and Hon Cheung Lee
- Subjects
Cyclic compounds -- Research ,Nucleotides -- Research ,Biological sciences ,Chemistry - Abstract
The cyclic nucleotide ADP-ribose, which is synthesized from NAD+ by ADP-ribosyl cyclase, increases the sensitivity of the Ca2+-releasing mechanism in various cells. It was shown that the ADP-ribosyl cyclase can also catalyze the cyclization of NAD+ with its purine bases to produce fluorescent analogs of ADP-ribose. The cyclization of the fluorescent analogs are at the N7 position of the purine ring rather than at the N1 position as in ADP-ribose. The variability of cyclization could affect the catalytic mechanism of ADP-ribose by altering its active-site structure.
- Published
- 1996
24. Calcium mobilization by dual receptors during fertilization of sea urchin eggs
- Author
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Hon Cheung Lee, Aarhus, Robert, and Walseth, Timothy F.
- Subjects
Sea urchins -- Eggs ,Cellular signal transduction -- Research ,Fertilization (Biology) -- Research - Published
- 1993
25. Porcine CD38 exhibits prominent secondary NAD+cyclase activity
- Author
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Kai Yiu Ting, Masayo Kotaka, Hon Cheung Lee, Quan Hao, Christina F. P. Leung, and Richard M. Graeff
- Subjects
0301 basic medicine ,Nicotinamide ,biology ,Chemistry ,Stereochemistry ,Active site ,Biochemistry ,Cyclic ADP-ribose ,Cyclase ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,Glycerol-3-phosphate dehydrogenase ,Hydrolase ,biology.protein ,NAD+ kinase ,Molecular Biology ,Cyclase activity - Abstract
Cyclic ADP-ribose (cADPR) mobilizes intracellular Ca(2+) stores and activates Ca(2+) influx to regulate a wide range of physiological processes. It is one of the products produced from the catalysis of NAD(+) by the multifunctional CD38/ADP-ribosyl cyclase superfamily. After elimination of the nicotinamide ring by the enzyme, the reaction intermediate of NAD(+) can either be hydrolyzed to form linear ADPR or cyclized to form cADPR. We have previously shown that human CD38 exhibits a higher preference towards the hydrolysis of NAD(+) to form linear ADPR while Aplysia ADP-ribosyl cyclase prefers cyclizing NAD(+) to form cADPR. In this study, we characterized the enzymatic properties of porcine CD38 and revealed that it has a prominent secondary NAD(+) cyclase activity producing cADPR. We also determined the X-ray crystallographic structures of porcine CD38 and were able to observe conformational flexibility at the base of the active site of the enzyme which allow the NAD(+) reaction intermediate to adopt conformations resulting in both hydrolysis and cyclization forming linear ADPR and cADPR respectively.
- Published
- 2016
26. 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
27. 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
28. 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
29. 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
30. Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38
- Author
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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
31. Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo
- Author
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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
32. 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
33. A Special Issue on Third and Fourth NAD (ASIAN 3) Meetings in 2014
- Author
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Antony Galione, Uh-Hyun Kim, Hon Cheung Lee, and Haruhiro Higashida
- Subjects
Chemistry ,Materials Science (miscellaneous) - Published
- 2014
34. Autocrine/Paracrine Function of Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) for Glucose Homeostasis in Pancreatic β-Cells and Adipocytes
- Author
-
Hon Cheung Lee, Asif Iqbal Shawl, Kwang-Hyun Park, Uh-Hyun Kim, Myung-Kwan Han, and Byung-Ju Kim
- Subjects
Male ,Biological Transport, Active ,Biology ,Second Messenger Systems ,Biochemistry ,Mice ,chemistry.chemical_compound ,Paracrine signalling ,Insulin-Secreting Cells ,Paracrine Communication ,Adipocytes ,medicine ,Animals ,Homeostasis ,Insulin ,Glucose homeostasis ,Calcium Signaling ,skin and connective tissue diseases ,Molecular Biology ,Nicotinic acid adenine dinucleotide phosphate ,Pancreatic islets ,Nucleoside inhibitor ,Cell Biology ,Autocrine Communication ,Kinetics ,Glucose ,medicine.anatomical_structure ,Diabetes Mellitus, Type 2 ,chemistry ,Second messenger system ,sense organs ,Beta cell ,NADP ,Signal Transduction - Abstract
Nicotinic acid adenine dinucleotide phosphate (NAADP) is a second messenger for mobilizing Ca(2+) from intracellular stores in various cell types. Extracellular application of NAADP has been shown to elicit intracellular Ca(2+) signals, indicating that it is readily transported into cells. However, little is known about the functional role of this NAADP uptake system. Here, we show that NAADP is effectively transported into selected cell types involved in glucose homeostasis, such as adipocytes and pancreatic β-cells, but not the acinar cells, in a high glucose-dependent manner. NAADP uptake was inhibitable by Ned-19, a NAADP mimic; dipyridamole, a nucleoside inhibitor; or NaN3, a metabolic inhibitor or under Ca(2+)-free conditions. Furthermore, NAADP was found to be released from pancreatic islets upon stimulation by high glucose. Consistently, administration of NAADP to type 2 diabetic mice improved glucose tolerance. We propose that NAADP is functioning as an autocrine/paracrine hormone important in glucose homeostasis. NAADP is thus a potential antidiabetic agent with therapeutic relevance.
- Published
- 2013
35. 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
36. An Improved Enzymatic Cycling Assay for NAADP
- Author
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Richard M. Graeff and Hon Cheung Lee
- Subjects
chemistry.chemical_classification ,Enzyme ,chemistry ,Biochemistry ,Materials Science (miscellaneous) ,Cycling - Published
- 2013
37. Poster session 6
- Author
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RM Melduni, Pittelkow, Shjj Hai, Kent R. Bailey, Bernard J. Gersh, SS Cha, and Hon Cheung Lee
- Subjects
medicine.medical_specialty ,Increased risk ,business.industry ,Physiology (medical) ,Internal medicine ,medicine ,Case-control study ,Cardiology ,Atrial fibrillation ,Dermatological disorders ,Cardiology and Cardiovascular Medicine ,medicine.disease ,business - Published
- 2013
38. Crystal Structures of Human CD38 in Complex with NAADP and ADPRP
- Author
-
Hongmin Zhang, Richard M. Graeff, Quan Hao, and Hon Cheung Lee
- Subjects
Chemistry ,Stereochemistry ,Materials Science (miscellaneous) ,Hydrolase ,Crystal structure ,CD38 - Published
- 2013
39. Dopamine-Induced Regulation and Deregulation of the Catabolism of Cyclic ADP-Ribose, an Intrinsic mTOR Signal Inhibitor, During Development in the Rodent Striatum
- Author
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Haruhiro Higashida, Chiharu Higashida, Mohammad Saharul Islam, Minako Hashii, Shin-ya Kamimura, Hon Cheung Lee, Jing Zhong, Duo Jin, Mingkun Liang, Richard M. Graeff, Osamu Hori, Hiroshi Okamoto, Jiasheng Zhang, Kohji Fukunaga, and Takeshi Inoue
- Subjects
Rodent ,biology ,Catabolism ,Chemistry ,Materials Science (miscellaneous) ,Striatum ,Cyclic ADP-ribose ,Cell biology ,chemistry.chemical_compound ,Dopamine ,biology.animal ,medicine ,PI3K/AKT/mTOR pathway ,medicine.drug - Published
- 2013
40. Intracellular Calcium Concentrations Regulated by Cyclic ADP-Ribose and Heat in the Mouse Hypothalamus
- Author
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Hon Cheung Lee, Mohammad Saharul Islam, Sarwat Amina, Minako Hashii, Richard M. Graeff, Olga Lopatina, Haruhiro Higashida, Chiharu Higashida, and Hong-Xiang Liu
- Subjects
chemistry.chemical_compound ,chemistry ,Materials Science (miscellaneous) ,Mouse Hypothalamus ,Cyclic ADP-ribose ,Calcium in biology ,Cell biology - Published
- 2012
41. Cyclic ADP-Ribose and Heat Regulate Oxytocin Release via CD38 and TRPM2 in the Hypothalamus during Social or Psychological Stress in Mice
- Author
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Shirin Akther, Haruhiro Higashida, Teruko Yuhi, Tomoko Nishimura, Minako Hashii, Makoto Tominaga, Shigeru Yokoyama, Olga Lopatina, Yasuhiko Yamamoto, Takahiro Tsuji, Yong Juan Zhao, Hiroshi Okamoto, Sarwat Amina, Hong-Xiang Liu, Hon Cheung Lee, Mingkun Liang, Kazumi Furuhara, Chiharu Tsuji, and Jing Zhong
- Subjects
0301 basic medicine ,Hyperthermia ,medicine.medical_specialty ,autism ,Stimulation ,Biology ,Cyclic ADP-ribose ,lcsh:RC321-571 ,03 medical and health sciences ,chemistry.chemical_compound ,stress ,0302 clinical medicine ,Endocrinology ,Internal medicine ,oxytocin ,Extracellular ,medicine ,cyclic ADP-ribose ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,Original Research ,Ryanodine receptor ,General Neuroscience ,medicine.disease ,hyperthermia ,NAD ,secretion ,030104 developmental biology ,Oxytocin ,chemistry ,Hypothalamus ,Knockout mouse ,030217 neurology & neurosurgery ,medicine.drug - 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 β-NAD+, cADPR, and ADP in mouse oxytocinergic neurons. As these β-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 min 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. © 2016 Zhong, Amina, Liang, Akther, Yuhi, Nishimura, Tsuji, Tsuji, Liu, Hashii, Furuhara, Yokoyama, Yamamoto, Okamoto, Zhao, Lee, Tominaga, Lopatina and Higashida.
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- 2016
- Full Text
- View/download PDF
42. Immuno-targeting the multifunctional CD38 using nanobody
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Shali Qi, Xian Wang Wang, Friedrich Koch-Nolte, Jun Liu, Mandy Unger, Du Xin, Quan Hao, Connie Mo Ching Lam, Hongmin Zhang, Yong Juan Zhao, Qi Wen Deng, Hon Cheung Lee, Yun Nan Hou, Ting Li, and Peng Zhang
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Models, Molecular ,0301 basic medicine ,medicine.medical_treatment ,Cell ,Biology ,CD38 ,Crystallography, X-Ray ,Article ,Epitope ,Cell Line ,03 medical and health sciences ,Antineoplastic Agents, Immunological ,0302 clinical medicine ,Antigen ,Antibody Specificity ,immune system diseases ,Immunotoxin ,Cell Line, Tumor ,hemic and lymphatic diseases ,medicine ,Humans ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Cytotoxicity ,Membrane Glycoproteins ,Multidisciplinary ,Immunotherapy ,Single-Domain Antibodies ,ADP-ribosyl Cyclase 1 ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,Cell culture ,030220 oncology & carcinogenesis ,Protein Conformation, beta-Strand ,Drug Screening Assays, Antitumor ,Multiple Myeloma ,Protein Binding - Abstract
CD38, as a cell surface antigen is highly expressed in several hematologic malignancies including multiple myeloma (MM) and has been proven to be a good target for immunotherapy of the disease. CD38 is also a signaling enzyme responsible for the metabolism of two novel calcium messenger molecules. To be able to target this multifunctional protein, we generated a series of nanobodies against CD38 with high affinities. Crystal structures of the complexes of CD38 with the nanobodies were solved, identifying three separate epitopes on the carboxyl domain. Chromobodies, engineered by tagging the nanobody with fluorescence proteins, provide fast, simple and versatile tools for quantifying CD38 expression. Results confirmed that CD38 was highly expressed in malignant MM cells compared with normal white blood cells. The immunotoxin constructed by splicing the nanobody with a bacterial toxin, PE38 shows highly selective cytotoxicity against patient-derived MM cells as well as the cell lines, with half maximal effective concentration reaching as low as 10−11 molar. The effectiveness of the immunotoxin can be further increased by stimulating CD38 expression using retinoid acid. These results set the stage for the development of clinical therapeutics as well as diagnostic screening for myeloma.
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- 2016
- Full Text
- View/download PDF
43. Asian-Pacific clinical practice guidelines on the management of hepatitis B: a 2015 update
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Wasim Jafri, Rosmawati Mohamed, Jidong Jia, Man-Fung Yuen, Jia-Horng Kao, Edward Gane, Stephen Locarnini, Pei-Jer Chen, Abdul Kadir Dokmeci, Hon Cheung Lee, M. Al Mahtab, Huey-Ling Chen, Jun Yong Park, Barjesh Chander Sharma, Jose D. Sollano, George K. K. Lau, Fu-Sheng Wang, J. H. Kim, Chien-Jen Chen, Teerha Piratvisuth, Manoj Kumar, Henry Lik-Yuen Chan, Seng Gee Lim, Ding-Shinn Chen, Ching-Lung Lai, Shusen Zheng, J L Hou, Rong-Nan Chien, Chun-Jen Liu, Shiv Kumar Sarin, Zaigham Abbas, Masao Omata, and Lai Wei
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Male ,Hepatitis B virus ,medicine.medical_specialty ,Asia ,Alternative medicine ,Disease ,Guidelines ,Bioinformatics ,Antiviral Agents ,03 medical and health sciences ,Hepatitis B, Chronic ,0302 clinical medicine ,Internal medicine ,HBV ,medicine ,Humans ,Acute hepatitis ,Hepatology ,business.industry ,Public health ,Disease Management ,Guideline ,Hepatitis B ,medicine.disease ,Vaccination ,Natural history ,030220 oncology & carcinogenesis ,Family medicine ,Acute Disease ,Africa ,Female ,030211 gastroenterology & hepatology ,business - Abstract
Worldwide, some 240 million people have chronic hepatitis B virus (HBV), with the highest rates of infection in Africa and Asia. Our understanding of the natural history of HBV infection and the potential for therapy of the resultant disease is continuously improving. New data have become available since the previous APASL guidelines for management of HBV infection were published in 2012. The objective of this manuscript is to update the recommendations for the optimal management of chronic HBV infection. The 2015 guidelines were developed by a panel of Asian experts chosen by the APASL. The clinical practice guidelines are based on evidence from existing publications or, if evidence was unavailable, on the experts' personal experience and opinion after deliberations. Manuscripts and abstracts of important meetings published through January 2015 have been evaluated. This guideline covers the full spectrum of care of patients infected with hepatitis B, including new terminology, natural history, screening, vaccination, counseling, diagnosis, assessment of the stage of liver disease, the indications, timing, choice and duration of single or combination of antiviral drugs, screening for HCC, management in special situations like childhood, pregnancy, coinfections, renal impairment and pre- and post-liver transplant, and policy guidelines. However, areas of uncertainty still exist, and clinicians, patients, and public health authorities must therefore continue to make choices on the basis of the evolving evidence. The final clinical practice guidelines and recommendations are presented here, along with the relevant background information.
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- 2016
44. Mechanism-based small molecule probes for labeling CD38 on live cells
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Hong Jiang, Congleton, Johanna, Qun Liu, Merchant, Paolomi, Malavasi, Fabio, Quan Hao, Hon Cheung Lee, Yen, Andrew, and Hening Lin
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Calcium -- Chemical properties ,Calcium -- Atomic properties ,Cytokines -- Chemical properties ,Membrane proteins -- Chemical properties ,NAD (Coenzyme) -- Chemical properties ,Chemistry - Abstract
The covalent trapping of mammalian CD38, a type II transmembrane protein, by arabinosyl-2'-fluoro-2'-deoxy (F-araNAD) is used to develop a mechanism-based labeling method. The labeling method could be used to study the CD38-mediated signaling processes and label other cell surface proteins.
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- 2009
45. A Novel Fluorescent Cell Membrane-permeable Caged Cyclic ADP-ribose Analogue
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Liangren Zhang, Baixia Hao, Yong Juan Zhao, Lihe Zhang, Jianbo Yue, Peilin Yu, Zhe-Hao Zhang, and Hon Cheung Lee
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Cell Membrane - metabolism ,Cyclic ADP-Ribose - analogs and derivatives - metabolism ,Blotting, Western ,TRPM Cation Channels ,Alkenes ,Biology ,Real-Time Polymerase Chain Reaction ,Biochemistry ,Jurkat cells ,Cyclic ADP-ribose ,Fluorescence ,Cell Line ,Cell membrane ,Jurkat Cells ,chemistry.chemical_compound ,Coumarins - metabolism ,Coumarins ,medicine ,Humans ,Alkenes - metabolism ,TRPM2 ,Calcium Signaling ,TRP Channels ,Stromal Interaction Molecule 1 ,Inosine ,Molecular Biology ,Cyclic ADP-Ribose ,Ryanodine receptor ,Endoplasmic reticulum ,Cell Membrane ,Membrane Proteins ,Ryanodine Receptor Calcium Release Channel ,STIM1 ,Cell Biology ,Neoplasm Proteins ,Cell biology ,HEK293 Cells ,medicine.anatomical_structure ,chemistry ,Calcium ,Ryanodine Receptor ,Signal Transduction ,SOCE ,medicine.drug - Abstract
Cyclic adenosine diphosphate ribose is an endogenous Ca(2+) mobilizer involved in diverse cellular processes. A cell membrane-permeable cyclic adenosine diphosphate ribose analogue, cyclic inosine diphosphoribose ether (cIDPRE), can induce Ca(2+) increase in intact human Jurkat T-lymphocytes. Here we synthesized a coumarin-caged analogue of cIDPRE (Co-i-cIDPRE), aiming to have a precisely temporal and spatial control of bioactive cIDPRE release inside the cell using UV uncaging. We showed that Co-i-cIDPRE accumulated inside Jurkat cells quickly and efficiently. Uncaging of Co-i-cIDPRE evoked Ca(2+) release from endoplasmic reticulum, with concomitant Ca(2+) influx in Jurkat cells. Ca(2+) release evoked by uncaged Co-i-cIDPRE was blocked by knockdown of ryanodine receptors (RyRs) 2 and 3 in Jurkat cells. The associated Ca(2+) influx, on the other hand, was abolished by double knockdown of Stim1 and TRPM2 in Jurkat cells. Furthermore, Ca(2+) release or influx evoked by uncaged Co-i-cIDPRE was recapitulated in HEK293 cells that overexpress RyRs or TRPM2, respectively, but not in wild-type cells lacking these channels. In summary, our results indicate that uncaging of Co-i-cIDPRE incites Ca(2+) release from endoplasmic reticulum via RyRs and triggers Ca(2+) influx via TRPM2., link_to_OA_fulltext
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- 2012
46. MESSENGER The Cyclic ADP-Ribose/NAADP/CD38-Signaling Pathway: Past and Present
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Hon Cheung Lee
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chemistry.chemical_compound ,Chemistry ,Materials Science (miscellaneous) ,CD38 ,Signal transduction ,Cyclic ADP-ribose ,Cell biology - Published
- 2012
47. Design, Synthesis and Biological Evaluation of Noncovalent Inhibitors of Human CD38 NADase
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Zhenming Liu, Jie Xia, Lihe Zhang, Liangren Zhang, Anna Ka Yee Kwong, Yi Zhou, Kai Yiu Ting, Hongwei Jin, Hon Cheung Lee, and Connie Mo Ching Lam
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Pharmacology ,Binding Sites ,Chemistry ,Phenylurea Compounds ,Organic Chemistry ,ADP-ribosyl Cyclase 1 ,Biochemistry ,Enzyme Activation ,Structure-Activity Relationship ,Design synthesis ,Catalytic Domain ,Drug Design ,Drug Discovery ,Humans ,Molecular Medicine ,Structure–activity relationship ,Enzyme Inhibitors ,General Pharmacology, Toxicology and Pharmaceutics ,Binding site ,Furans ,Biological evaluation - Published
- 2012
48. [The synthesis of purine derivatives and its inhibitory activity on CD38 NADase]
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Na, Li, Wen-jie, Zhu, Xi-wen, Xue, Yong-juan, Zhao, Hon-cheung, Lee, Liang-ren, Zhang, and Li-he, Zhang
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Purines ,Humans ,Enzyme Inhibitors ,ADP-ribosyl Cyclase 1 - Abstract
CD38 is a multifunctional enzyme expressed in a variety of mammalian tissues, its catalytic activity was involved in a wide range of physiological processes. Based on the reported inhibitor of human CD38 NADase, 33 purine derivatives were designed and synthesized. The biological activity assay showed that compounds 20 and 38 exhibited almost the same extent of inhibitory activities on human CD38 NADase as the lead compound H2. The results also revealed that small substituents at C-6 of purine ring gave no obvious effect on inhibitory activity, but phenylpropionyl moiety at N-2 could affect the binding mode of the compound with CD38. This study provides a reliable basis for future rational design of inhibitors for CD38.
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- 2015
49. Conformational closure of the catalytic site of human CD38 induced by calcium
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Qun Liu, Graeff, Richard, Kriksunov, Irina A., Lam, Connie M.C., Hon Cheung Lee, and Quan Hao
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Calcium-binding proteins -- Structure ,Calcium-binding proteins -- Chemical properties ,Catalysis -- Analysis ,Hydrolysis -- Analysis ,Hydrophobic effect -- Analysis ,Biological sciences ,Chemistry - Abstract
The crystal structure of the calcium-loaded human CD38 at 1.45 [Angstrom] resolutions is determined, which has disclosed that CD38 has undergone dramatic structural changes to an inhibited conformation in the presence of calcium. The results have shown a novel conformational coupling mechanism, whereby protein interaction targeted at the receptor region has effectively regulated activity of CD38.
- Published
- 2008
50. Hierarchical and helical self-assembly of ADP-ribosyl cyclase into large-scale protein microtubes
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Qun Liu, Kriksunov, Irina A., Zhongwu Wang, Graeff, Richard, Hon Cheung Lee, and Quan Hao
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Scanning electron microscopes -- Usage ,X-ray crystallography -- Usage ,Proteins -- Structure ,Proteins -- Analysis ,Chemicals, plastics and rubber industries - Published
- 2008
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