DICP OpenIR
Trypsin-Catalyzed N-Terminal Labeling of Peptides with Stable Isotope-Coded Affinity Tags for Proteome Analysis
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
Source PublicationANALYTICAL CHEMISTRY
2014-01-21
DOI10.1021/ac403060d
Volume86Issue:2Pages:1170-1177
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordMASS-SPECTROMETRY ; QUANTITATIVE PROTEOMICS ; SULFONATED PEPTIDES ; ISOBARIC TAGS ; QUANTIFICATION ; GUANIDINATION ; DISSOCIATION ; FRAGMENTS
AbstractAn 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.
Language英语
WOS IDWOS:000330204500029
Citation statistics
Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/145495
Collection中国科学院大连化学物理研究所
Affiliation1.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
GB/T 7714
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.
APA Pan, Yanbo.,Ye, Mingliang.,Zheng, Hao.,Cheng, Kai.,Sun, Zhen.,...&Zou, Hanfa.(2014).Trypsin-Catalyzed N-Terminal Labeling of Peptides with Stable Isotope-Coded Affinity Tags for Proteome Analysis.ANALYTICAL CHEMISTRY,86(2),1170-1177.
MLA Pan, Yanbo,et al."Trypsin-Catalyzed N-Terminal Labeling of Peptides with Stable Isotope-Coded Affinity Tags for Proteome Analysis".ANALYTICAL CHEMISTRY 86.2(2014):1170-1177.
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