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  2. 上海药物研究所
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Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells

文献类型：期刊论文

作者Hu, Wei2; Yuan, Yi3; Wang, Cai-Hu2; Tian, Hong-Tao2,4; Guo, An-Di2,4; Nie, Hui-Jun2; Hu, Hao1; Tan, Minjia1; Tang, Zhuo3; Chen, Xiao-Hua2
刊名CHEM
出版日期2019-11-14
卷号5期号:11页码:2955-2968
ISSN号2451-9294
DOI10.1016/j.chempr.2019.08.020
通讯作者Tang, Zhuo(tangzhuo@cib.ac.cn) ; Chen, Xiao-Hua(xhchen@simm.ac.cn)
英文摘要The genetically encoded non-selective photo-crosslinkers suffer from unpredictable crosslinking sites and false positive interacting proteins, thus, the identification of protein interactions and mapping accurate interaction interfaces remain largely elusive in living systems. Here, we report genetically encoded, residue-selective protein photo-crosslinker enabling covalent crosslinking of the interacting proteins with proximal lysine, upon UV light activation in vitro and in living cells. Using this photo-crosslinker, we demonstrate a crosslinking strategy to capture protein-protein interactions with residue selectivity in a temporal-resolved manner. In addition, this crosslinking strategy produces the lysine-selective crosslinking site and predictable crosslinked peptide, which serve as the direct evidence for protein-protein interactions and facilitate mass spectrometry analysis. More importantly, this photo-crosslinker enables the capture of elusive enzyme-substrate interaction with directly interacting lysine and validation of acetylation site of the substrate. This strategy represents higher spatiotemporal resolution, accuracy, and reliability for investigating dynamic, transient, and weak protein-protein interactions in living systems.
WOS关键词LINKING MASS-SPECTROMETRY ; TRANSFER-RNA SYNTHETASE ; AMINO-ACID ; POSTTRANSLATIONAL MODIFICATIONS ; MAMMALIAN-CELLS ; CHEMISTRY ; CODE ; BINDING ; ENZYME ; SITE
资助项目National Science Foundation of China (NSFC)[21602235] ; National Science Foundation of China (NSFC)[21778062] ; Chinese Academy of Science ; Natural Science Foundation of Shanghai[16ZR1442800] ; Shanghai Rising-Star program
WOS研究方向Chemistry
语种英语
出版者CELL PRESS
WOS记录号WOS:000496957600019
源URL[http://119.78.100.183/handle/2S10ELR8/282023]  
专题新药研究国家重点实验室
通讯作者Tang, Zhuo; Chen, Xiao-Hua
作者单位1.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China
2.Chinese Acad Sci, Key Lab Receptor Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
3.Chinese Acad Sci, Nat Prod Res Ctr, Chengdu Inst Biol, Chengdu 610041, Sichuan, Peoples R China
4.Univ Chinese Acad Sci, 19A Yuquan Rd, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714		 Hu, Wei,Yuan, Yi,Wang, Cai-Hu,et al. Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells[J]. CHEM,2019,5(11):2955-2968.
APA Hu, Wei.,Yuan, Yi.,Wang, Cai-Hu.,Tian, Hong-Tao.,Guo, An-Di.,...&Chen, Xiao-Hua.(2019).Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells.CHEM,5(11),2955-2968.
MLA Hu, Wei,et al."Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells".CHEM 5.11(2019):2955-2968.

入库方式： OAI收割

来源：上海药物研究所

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