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Tryptic Peptides Bearing C-Terminal Dimethyllysine Need to Be Considered during the Analysis of Lysine Dimethylation in Proteomic Study

文献类型：期刊论文

作者Chen, Ming1,2,3; Zhang, Min1,2,3; Zhai, Linhui1,3; Hu, Hao1,3; Liu, Ping1,3; Tan, Minjia1,2,3
刊名JOURNAL OF PROTEOME RESEARCH
出版日期2017-09
卷号16期号:9页码:3460-3469
关键词trypsin lysine dimethylation proteomics terminal amine isotopic labeling of substrates (TAILS) mass spectrometry analysis
ISSN号1535-3893
DOI10.1021/acs.jproteome.7b00373
文献子类Article
英文摘要Lysine methylation plays important roles in structural and functional regulation of chromatin. Although trypsin is the most widely used protease in mass spectrometry-based proteomic analysis for lysine methylation substrates, the proteolytic activity of trypsin on dimethylated lysine residues remains an arguable issue. In this study, we tested the ability of trypsin to cleave dimethylated lysine residues in synthetic peptides, purified albumin, and whole cell lysate, and found that the C-terminal of dimethylated lysine residue could be cleaved in a protein sequence dependent manner. Kinetic studies revealed that the optimal digestion time and enzyme-to-substrate ratio for the cleavage of dimethylated lysine by trypsin was around 16 h and 1:50, respectively. We further showed the tryptic C-terminal lysine-dimethylated (C-Kme2) peptides could contribute to a significant portion of substrate identification in the proteomic study, which utilizes the chemical dimethylation labeling approach. More than 120 tryptic C-Kme2 peptides (7% of total peptides identified) were identified in chemically lysine-dimethyl-labeled HeLa whole cell lysate by a single-shot nanoflow high performance liquid chromatography with tandem mass spectrometry (nano-HPLC-MS/MS) analysis. Moreover, in an assay for substrate identification of protease Glu-C using stable isotope dimethyl labeling approach, our data showed the tryptic C-Kme2 peptides accounted for more than 13% of total tryptic peptides. Additionally, our in vivo methylome profiling data revealed some C-Kme2 peptides, which is of importance to identification and quantification of biologically relevant protein and lysine-methylated site. Therefore, we reason that the tryptic peptides bearing C-terminal dimethylated lysine need to be considered in the mass spectrometric analysis of lysine dimethylation.
WOS关键词POSTTRANSLATIONAL MODIFICATIONS ; MASS-SPECTROMETRY ; STEM-CELLS ; N-TERMINI ; PROTEINS ; IDENTIFICATION ; METHYLATION ; TRYPSIN ; METHYLTRANSFERASES ; QUANTIFICATION
资助项目National Basic Research Program of China (973 Program)[2014CBA02001] ; Natural Science Foundation of China[31670066] ; Youth Science and Technology Talents in Shanghai Sail Plan of China[16YF1414000] ; Youth Science and Technology Talents in Shanghai Sail Plan of China[17YF1423200] ; Chinese Academy of Sciences[XDA12020314] ; Chinese Academy of Sciences[2060499]
WOS研究方向Biochemistry & Molecular Biology
语种英语
WOS记录号WOS:000410003500031
出版者AMER CHEMICAL SOC
源URL[http://119.78.100.183/handle/2S10ELR8/272505]  
专题化学蛋白质组学研究中心
中科院受体结构与功能重点实验室
新药研究国家重点实验室
通讯作者Tan, Minjia
作者单位1.Chinese Acad Sci, State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China;
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Chinese Acad Sci, Chem Prote Ctr, Shanghai 201203, Peoples R China;
推荐引用方式
GB/T 7714		 Chen, Ming,Zhang, Min,Zhai, Linhui,et al. Tryptic Peptides Bearing C-Terminal Dimethyllysine Need to Be Considered during the Analysis of Lysine Dimethylation in Proteomic Study[J]. JOURNAL OF PROTEOME RESEARCH,2017,16(9):3460-3469.
APA Chen, Ming,Zhang, Min,Zhai, Linhui,Hu, Hao,Liu, Ping,&Tan, Minjia.(2017).Tryptic Peptides Bearing C-Terminal Dimethyllysine Need to Be Considered during the Analysis of Lysine Dimethylation in Proteomic Study.JOURNAL OF PROTEOME RESEARCH,16(9),3460-3469.
MLA Chen, Ming,et al."Tryptic Peptides Bearing C-Terminal Dimethyllysine Need to Be Considered during the Analysis of Lysine Dimethylation in Proteomic Study".JOURNAL OF PROTEOME RESEARCH 16.9(2017):3460-3469.

入库方式： OAI收割

来源：上海药物研究所

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