文献类型：期刊文献

英文题名：Selective enrichment of N-terminal proline peptides via hydrazide chemistry for proteomics analysis

作者：Li, Zhouxian[1,2];Wang, Qi[2,3];Mao, Jiawei[2,3];Zhang, Lingyi[1];Zhang, Weibing[1];Ye, Mingliang[2,3]

机构：[1]East China Univ Sci & Technol, Shanghai Key Lab Funct Mat Chem, Dept Chem & Mol Engn, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China;[3]Univ Chinese Acad Sci, Beijing 100049, Peoples R China

年份：2021

卷号：1142

起止页码：48

外文期刊名：ANALYTICA CHIMICA ACTA

收录：;EI(收录号：20204509466584);WOS:【SCI-EXPANDED(收录号:WOS:000608817100006)】；

基金：This work was supported, in part, by funds from Science and Technology Commission of Shanghai Municipality (19142201100), the National Key Research and Development Program of China (2016YFA0501402, 2017YFA0505004), the National Natural Science Foundation of China (21974045, 21675061, 21535008, 91753105), the Fundamental Research Funds for the Central Universities (50321102017022), the LiaoNing Revitalization Talents Program, the innovation program (DICP I201935, DICP&QIBEBT UN201802) of science and research from the DICP, CAS, and Innovation Academy for Precision Measurement Science and Technology, CAS. M.Y. is a recipient of the National Science Fund of China for Distinguished Young Scholars (21525524).

语种：英文

外文关键词：N-terminal proline peptides; OPA blocking; Reductive glutaraldehydation; Solid phase hydrazide chemistry

摘要：A challenge for shotgun proteomics is the identification of low abundance proteins, which is always hampered owing to the extreme complexity of protein digests and highly dynamic concentration range of proteins. To reduce the complexity of the peptide mixture, we developed a novel method to selectively enrich N-terminal proline peptides via hydrazide chemistry. This method consisted of ortho-phthalaldehyde (OPA) blocking of primary amines in peptides, reductive glutaraldehydation of N-terminal proline and solid phase hydrazide chemistry enrichment of aldehyde-modified N-terminal proline peptide. After enrichment, the number of detected peptides containing N-terminal proline increased from 1304 to 4039 and the ratio of N-terminal proline peptides jumped from 4.4% to 93.7%, showing good enrichment specificity towards N-terminal proline peptides. Besides, the ratio of identified peptides to proteins was decreased from 7.8 (29751/3811) to 1.5 (4347/2821), indicating that sample complexity was drastically reduced through this method. As a result, this novel approach for enriching N-terminal proline peptides is effective in identification of low abundance protein owing to the reduction of sample complexity. (C) 2020 Elsevier B.V. All rights reserved.