文献类型：期刊文献

英文题名：Highly Efficient Separation of Methylated Peptides Utilizing Selective Complexation between Lysine and 18-Crown-6

作者：Sheng, Qianying[1];Wang, Cunli[2];Li, Xiaopei[2];Qin, Hongqiang[2];Ye, Mingliang[2];Xiong, Yuting[2];Wang, Xue[2];Li, Xiuling[2];Lan, Minbo[1];Li, Junyan[3];Ke, Yanxiong[3];Qing, Guangyan[2];Liang, Xinmiao[2,3]

机构：[1]East China Univ Sci & Technol, Shanghai Key Lab Funct Mat Chem, Sch Chem & Mol Engn, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China;[3]East China Univ Sci & Technol, Engn Res Ctr Pharmaceut Proc Chem, Sch Pharm, Minist Educ, Shanghai 200237, Peoples R China

年份：2020

卷号：92

期号：23

起止页码：15663

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20204809542434);WOS:【SCI-EXPANDED(收录号:WOS:000596727600052)】；

基金：This work was supported by the National Natural Science Foundation of China (21775116, 21804041, 21922411, and 21934005), DICP Innovation Funding (DICP-RC201801), and LiaoNing Revitalization Talents Program (XLYC1802109).

语种：英文

外文关键词：Body fluids - Signal transduction - High performance liquid chromatography - Separation - Amino acids - Molecular biology - Quantum chemistry - Alkylation - Mass spectrometry - Methylation - Signal to noise ratio

摘要：Protein methylation is one of the most common and important post-translational modifications, and it plays vital roles in epigenetic regulation, signal transduction, and chromatin metabolism. However, due to the diversity of methylation forms, slight difference between methylated sites and nonmodified ones, and ultralow abundance, it is extraordinarily challenging to capture and separate methylated peptides from biological samples. Here, we introduce a simple and highly efficient method to separate methylated and nonmethylated peptides using 18-crown-6 as a mobile phase additive in high-performance liquid chromatography. Selective complexation between lysine and 18-crown-6 remarkably increases the retention of the peptides on a C18 stationary phase, leading to an excellent baseline separation between the lysine methylated and nonmethylated peptides. A possible binding mechanism is verified by nuclear magnetic resonance titration, biolayer interferometry technology, and quantum chemistry calculation. Through establishment of a simple enrichment methodology, a good selectivity is achieved and four methylated peptides with greatly improved signal-to-noise (S/N) ratios are successfully separated from a complex peptide sample containing 10-fold bovine serum albumin tryptic digests. By selecting rLys N as an enzyme to digest histone, methylation information in the histone could be well identified based on our enrichment method. This study will open an avenue and provide a novel insight for selective enrichment of lysine methylated peptides in post-translational modification proteomics.