文献类型：期刊文献

英文题名：An antibody-free enrichment approach enabled by reductive glutaraldehydation for monomethyllysine proteome analysis

作者：Li, Zhouxian[1,2];Wang, Qi[2];Wang, Keyun[2];Zhang, Weibing[1];Ye, Mingliang[2]

机构：[1]East China Univ Sci & Technol, Dept Chem & Mol Engn, Shanghai Key Lab Funct Mat Chem, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

年份：2023

卷号：23

期号：3-4

外文期刊名：PROTEOMICS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000796371300001)】；

基金：Science and Technology Commission of Shanghai Municipality, Grant/Award Number: 19142201100; National Key Research and Development Program of China, Grant/Award Numbers: 2017YFA0505004, 2020YFE0202200; National Natural Science Foundation of China, Grant/Award Numbers: 21974045, 21675061, 92153302; Fundamental Research Funds for the Central Universities, Grant/Award Number: JKJ01211718; DICP; CAS, Grant/Award Numbers: I201935, UN201802

语种：英文

外文关键词：hydrazide chemistry; protein lysine monomethylation; reductive glutaraldehydation

摘要：Protein lysine monomethylation is an important post-translational modification participated in regulating many biological processes. There is growing interest in identifying these methylation events. However, the introduction of one methyl group on lysine residues has negligible effect on changing the physical and chemical properties of proteins or peptides, making enriching and identifying monomethylated lysine (Kme1) proteins or peptides extraordinarily challenging. In this study, we proposed an antibody-free chemical proteomics approach to capture Kme1 peptides from complex protein digest. By exploiting reductive glutaraldehydation, 5-aldehyde-pentanyl modified Kme1 residues and piperidine modified primary amines were generated at the same time. The peptides with aldehyde modified Kme1 residues were then enriched by solid-phase hydrazide chemistry. This chemical proteomics approach was validated by using several synthetic peptides. It was demonstrated that it can enrich and detect Kme1 peptide from peptide mixture containing 5000-fold more bovine serum albumin tryptic digest. Besides, we extended our approach to profile Kme1 using heavy methyl stable isotope labeling by amino acids in cell culture (hmSILAC) labeled Jurkat T cells and Hela cells. Totally, 29 Kme1 sites on 25 proteins were identified with high confidence and 11 Kme1 sites were identified in both two types cells. This is the first antibody-free chemical proteomics approach to enrich Kme1 peptides from complex protein digest, and it provides a potential avenue for the analysis of methylome.