A rapid and easy protein N-terminal profiling strategy using (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl) phosphonium bromide ( TMPP) labeling and StageTip

doi:10.1002/pmic.201600481

	A rapid and easy protein N-terminal profiling strategy using (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl) phosphonium bromide ( TMPP) labeling and StageTip
	Li, Yanchang 1; Wang, Zhiqiang 1,2,3; Zhou, Wenjing 4; Zhang, Kun 4; Ma, Jia 1; Wu, Feilin 1; Ji, Jianguo 5; Hong, Xuechuan 2,3; Deng, Zixin 2,3; He, Simin 4; Xu, Ping 1,2,3,6
	2017-07-01
发表期刊	PROTEOMICS
ISSN	1615-9853
卷号	17 期号:13-14 页码:6
摘要	Protein N-terminal profiling is crucial when characterizing biological functions and provides proteomic evidences for genome reannotations. However, most of the current N-terminal enrichment approaches involve multiple chemical derivatizations and chromatographic separation processes which are time consuming and can contribute to N-terminal peptide losses. In this study, a fast, one-step approach utilizing (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP) derivatization and StageTip separation was developed to enhance N-terminal peptide enrichment and analysis. Based on the characteristics of TMPP-derivatized samples, such as a higher hydrophobicity and increased likelihood to produce a and b ions in collision-induced dissociation or HCD fragmentation modes, first the SDS-PAGE was optimized to increase protein loading and gel entry and to remove unbound TMPP. Then, this process was combined with a simplified StageTip separation and a new scoring criterion (considering a, b and y ions) to identify more TMPP-modified N-terminal spectra. When utilizing a low amount of starting material (similar to 20 mu g protein), a total of 581 yeast N-terminal peptides were identified, with 485 of them being TMPP modified, in only about one third of the general experimental time. It is hoped that the workflow constructed herein will provide a fast and practical strategy for N-terminomic studies.
关键词	Protein N-termini Proteomics Spectrum scoring StageTip TMPP
DOI	10.1002/pmic.201600481
收录类别	SCI
语种	英语
资助项目	Chinese National Basic Research Programs[2016YFA0501300] ; Chinese National Basic Research Programs[2013CB911201] ; Chinese National Basic Research Programs[2015CB910700] ; National Natural Science Foundation of China[31470809] ; National Natural Science Foundation of China[31670834] ; National Natural Science Foundation of China[31400698] ; National Natural Science Foundation of China[31400697] ; International Collaboration Program[2014DFB30020] ; National Natural Science Foundation of Beijing[5152008] ; Beijing Training Project for The Leading Talents in S&T, National Megaprojects for Key Infectious Diseases[2013zx10003002] ; Foundation of State Key Lab of Proteomics[SKLP-Y201501] ; Innovation Foundation of Medicine[2017CXJJ19] ; Innovation Foundation of Medicine[16CXZ027] ; Innovation Foundation of Medicine[BWS14J052] ; Tianjin Baodi hospital Translational Medicine Research Center[TMRC2014M02B] ; Unilevel 21th Century Toxicity Program[MA-2014-02409]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology
WOS记录号	WOS:000405754500008
出版者	WILEY
引用统计	被引频次：11[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://119.78.100.204/handle/2XEOYT63/6919
专题	中国科学院计算技术研究所期刊论文_英文
通讯作者	Xu, Ping
作者单位	1.Beijing Inst Radiat Med, Beijing Proteome Res Ctr, State Key Lab Prote, Natl Ctr Prot Sci Beijing, Beijing, Peoples R China 2.Wuhan Univ, Sch Pharmaceut Sci, Key Lab Combinatorial Biosynthesis & Drug Discove, Wuhan, Peoples R China 3.Wuhan Univ, Sch Pharmaceut Sci, Hubei Prov Engn & Technol Res Ctr Fluorinated Pha, State Key Lab Virol, Wuhan, Peoples R China 4.Chinese Acad Sci, Inst Comp Technol, Key Lab Intelligent Informat Proc, Beijing, Peoples R China 5.Peking Univ, Coll Life Sci, State Key Lab Prot & Plant Gene Res, Beijing, Peoples R China 6.Anhui Med Univ, Hefei, Peoples R China
推荐引用方式 GB/T 7714	Li, Yanchang,Wang, Zhiqiang,Zhou, Wenjing,et al. A rapid and easy protein N-terminal profiling strategy using (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl) phosphonium bromide ( TMPP) labeling and StageTip[J]. PROTEOMICS,2017,17(13-14):6.
APA	Li, Yanchang.,Wang, Zhiqiang.,Zhou, Wenjing.,Zhang, Kun.,Ma, Jia.,...&Xu, Ping.(2017).A rapid and easy protein N-terminal profiling strategy using (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl) phosphonium bromide ( TMPP) labeling and StageTip.PROTEOMICS,17(13-14),6.
MLA	Li, Yanchang,et al."A rapid and easy protein N-terminal profiling strategy using (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl) phosphonium bromide ( TMPP) labeling and StageTip".PROTEOMICS 17.13-14(2017):6.