Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states

doi:10.7554/eLife.12509

CORC > 计算技术研究所 > 中国科学院计算技术研究所 > 中国科学院计算技术研究所期刊论文 > 英文

	Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states
	Tan, Dan 2,3; Li, Qiang 1,2,6,7; Zhang, Mei-Jun 2; Liu, Chao 5; Ma, Chengying 4; Zhang, Pan 2,3; Ding, Yue-He 2,3; Fan, Sheng-Bo 5; Tao, Li 2,3; Yang, Bing 2
刊名	ELIFE
	2016-03-08
卷号	5 页码:51
ISSN号	2050-084X
DOI	10.7554/eLife.12509
英文摘要	To improve chemical cross-linking of proteins coupled with mass spectrometry (CXMS), we developed a lysine-targeted enrichable cross-linker containing a biotin tag for affinity purification, a chemical cleavage site to separate cross-linked peptides away from biotin after enrichment, and a spacer arm that can be labeled with stable isotopes for quantitation. By locating the flexible proteins on the surface of 70S ribosome, we show that this trifunctional cross-linker is effective at attaining structural information not easily attainable by crystallography and electron microscopy. From a crude Rrp46 immunoprecipitate, it helped identify two direct binding partners of Rrp46 and 15 protein-protein interactions (PPIs) among the co-immunoprecipitated exosome subunits. Applying it to E. coli and C. elegans lysates, we identified 3130 and 893 inter-linked lysine pairs, representing 677 and 121 PPIs. Using a quantitative CXMS workflow we demonstrate that it can reveal changes in the reactivity of lysine residues due to protein-nucleic acid interaction.
资助项目	National Natural Science Foundation of China ; Ministry of Science and Technology of the People's Republic of China ; Chinese Academy of Sciences
WOS研究方向	Life Sciences & Biomedicine - Other Topics
语种	英语
出版者	ELIFE SCIENCES PUBLICATIONS LTD
WOS记录号	WOS:000373894300001
内容类型	期刊论文
源URL	[http://119.78.100.204/handle/2XEOYT63/8460]
专题	中国科学院计算技术研究所期刊论文_英文
通讯作者	Dong, Meng-Qiu; Lei, Xiaoguang
作者单位	1.Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China 2.Natl Inst Biol Sci, Beijing, Peoples R China 3.Chinese Acad Med Sci, Peking Union Med Coll, Grad Program, Beijing 100730, Peoples R China 4.Tsinghua Univ, Sch Life Sci, Key Lab Prot Sci, Minist Educ, Beijing 100084, Peoples R China 5.Chinese Acad Sci, Inst Comp Technol, Key Lab Intelligent Informat Proc Chinese Acad Sc, Beijing, Peoples R China 6.Peking Univ, Coll Chem & Mol Engn, Dept Biol Chem, Beijing 100871, Peoples R China 7.Peking Univ, Synthet & Funct Biomol Ctr, Beijing 100871, Peoples R China
推荐引用方式 GB/T 7714	Tan, Dan,Li, Qiang,Zhang, Mei-Jun,et al. Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states[J]. ELIFE,2016,5:51.
APA	Tan, Dan.,Li, Qiang.,Zhang, Mei-Jun.,Liu, Chao.,Ma, Chengying.,...&Lei, Xiaoguang.(2016).Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states.ELIFE,5,51.
MLA	Tan, Dan,et al."Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states".ELIFE 5(2016):51.