Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells

doi:10.1016/j.chempr.2019.08.020

	Genetically Encoded Residue-Selective Photo-Crosslinker to Capture Protein-Protein Interactions in Living Cells
	Hu, Wei 2; Yuan, Yi 3; Wang, Cai-Hu 2; Tian, Hong-Tao 2,4; Guo, An-Di 2,4; Nie, Hui-Jun 2; Hu, Hao 1; Tan, Minjia 1; Tang, Zhuo 3; Chen, Xiao-Hua 2
刊名	CHEM
	2019-11-14
卷号	5 期号:11 页码:2955-2968
ISSN号	2451-9294
DOI	10.1016/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.
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关键词	LINKING MASS-SPECTROMETRY ; TRANSFER-RNA SYNTHETASE ; AMINO-ACID ; POSTTRANSLATIONAL MODIFICATIONS ; MAMMALIAN-CELLS ; CHEMISTRY ; CODE ; BINDING ; ENZYME ; SITE
WOS研究方向	Chemistry
语种	英语
出版者	CELL PRESS
WOS记录号	WOS:000496957600019
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/282023]
专题	中国科学院上海药物研究所
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.