title: Identifying Glyceraldehyde 3-Phosphate Dehydrogenase as a Cyclic Adenosine Diphosphoribose Binding Protein by Photoaffinity Protein-Ligand Labeling Approach.
abstract: Cyclic adenosine diphosphoribose (cADPR), an endogenous nucleotide derived from nicotinamide adenine dinucleotide (NAD+), mobilizes Ca2+release from endoplasmic reticulum (ER) via ryanodine receptors (RyRs), yet the bridging protein(s) between cADPR and RyRs remain(s) unknown. Here we synthesized a novel photoaffinity labeling (PAL) cADPR agonist, PAL-cIDPRE, and subsequently applied it to purify its binding proteins in human Jurkat T cells. We identified glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as one of the cADPR binding protein(s), characterized the binding affinity between cADPR and GAPDH in vitro by surface plasmon resonance (SPR) assay, and mapped cADPR's binding sites in GAPDH. We further demonstrated that cADPR induces the transient interaction between GAPDH and RyRs in vivo and that GAPDH knockdown abolished cADPR-induced Ca2+release. However, GAPDH did not catalyze cADPR into any other known or novel compound(s). In summary, our data clearly indicate that GAPDH is the long-sought-after cADPR binding protein and is required for cADPR-mediated Ca2+mobilization from ER via RyRs.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University , Beijing 100191, China.|Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Kehui
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.|Department of Biology and Shenzhen Key Laboratory of Cell Microenvironment, South University of Science and Technology of China , Shenzhen 518052, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Kaiyuan
authors.initials: K
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University , Beijing 100191, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Pei
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Longchao
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Biology and Shenzhen Key Laboratory of Cell Microenvironment, South University of Science and Technology of China , Shenzhen 518052, China.
authors.forename: Hongmin
authors.initials: H
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University , Beijing 100191, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University , Beijing 100191, China.
authors.forename: Li-He
authors.initials: LH
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University , Beijing 100191, China.
authors.forename: Liangren
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong , Hong Kong, China.
authors.forename: Jianbo
authors.identifier: 0000-0001-6384-5447
authors.initials: J
authors.lastname: Yue
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000737:chemistry / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D019920:Photoaffinity Labels / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D008024:Ligands; D019920:Photoaffinity Labels; D000246:Adenosine Diphosphate Ribose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases