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题名: Trypsin-Catalyzed N-Terminal Labeling of Peptides with Stable Isotope-Coded Affinity Tags for Proteome Analysis
作者: Pan, Yanbo1, 2;  Ye, Mingliang1;  Zheng, Hao1, 2;  Cheng, Kai1, 2;  Sun, Zhen1, 2;  Liu, Fangjie1, 2;  Liu, Jing1, 2;  Wang, Keyun1, 2;  Qin, Hongqiang1;  Zou, Hanfa1
刊名: ANALYTICAL CHEMISTRY
发表日期: 2014-01-21
DOI: 10.1021/ac403060d
卷: 86, 期:2, 页:1170-1177
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: An enzymatic approach to label peptide N-termini with isotope-coded affinity tags is presented. This method exploits the high activity of trypsin for peptide synthesis in organic solvents. A cosubstrate containing a stable isotope-coded Arg residue and a biotin tag was synthesized. When the cosubstrate was incubated with tryptic peptides and trypsin in ethanol solution, the stable isotope-coded affinity tag was specifically coupled onto the N-termini of peptides via the formation of new peptide bonds. The labeled peptides were specifically enriched by avidin affinity chromatography and then were submitted to liquid chromatography tandem mass spectrometry (LC/MS/MS) for quantification. This enrichment step effectively reduced the interference by unlabeled peptides. The excellent performance of this approach was demonstrated by labeling standard peptides as well as a mouse liver digest. In addition to one amino acid residue, a few dipeptide tags were also introduced to the N-termini of peptides successfully by this enzymatic approach. It was found that the identifications for samples labeled with these tags were highly complementary. Coupling a short sequence tag onto peptides could be an effective approach to improve the coverage for proteome analysis.
关键词[WOS]: MASS-SPECTROMETRY ;  QUANTITATIVE PROTEOMICS ;  SULFONATED PEPTIDES ;  ISOBARIC TAGS ;  QUANTIFICATION ;  GUANIDINATION ;  DISSOCIATION ;  FRAGMENTS
语种: 英语
WOS记录号: WOS:000330204500029
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/145495
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

Recommended Citation:
Pan, Yanbo,Ye, Mingliang,Zheng, Hao,et al. Trypsin-Catalyzed N-Terminal Labeling of Peptides with Stable Isotope-Coded Affinity Tags for Proteome Analysis[J]. ANALYTICAL CHEMISTRY,2014,86(2):1170-1177.
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