Rapid and large-scale glycopeptide enrichment strategy based on chemical ligation  

作　　者：Yingying Xiong Zhuoer Lu Yuyin Shao Peiyi Meng Guoli Wang Xinwen Zhou Jun Yao Huimin Bao Haojie Lu 

机构地区：[1]Department of Chemistry and Liver Cancer Institute,Zhongshan Hospital,Fudan University,Shanghai 200032,China [2]Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research,Fudan University,Shanghai 200032,China

出　　处：《National Science Review》2024年第11期142-153,共12页国家科学评论（英文版）

基　　金：supported by the National Key Research and Development Program of China(2023YFF1205900);the National Natural Science Foundation of China(22434001 and 82121004);the Shanghai Science and Technology Program(2023SHZDZX02,22DZ2291700 and 22142202400).

摘　　要：Protein glycosylation,the most universal post-translational modification,is thought to play a crucial role in regulating multiple essential cellular processes.However,the low abundance of glycoproteins and the heterogeneity of glycans complicate their comprehensive analysis.Here,we develop a rapid and large-scale glycopeptide enrichment strategy via bioorthogonal ligation and trypsin cleavage.The enrichment process is performed in one tube to minimize sample loss and time costs.This method combines convenience and practicality,identifying over 900 O-GlcNAc sites from a 500μg sample.Surprisingly,it allows simultaneous identification of N-glycosites,O-GlcNAc sites,O-GalNAc sites and N-glycans via a two-step enzymatic release strategy.Combined with quantitative analysis,it reveals the distinct O-GlcNAcylation patterns in different compartments during oxidative stress.In summary,our study offers a convenient and robust tool for glycoproteome and glycome profiling,facilitating in-depth analysis to elucidate the biological functions of glycosylation.

关 键 词：GLYCOPROTEOMICS large-scale enrichment O-GLCNACYLATION N-GLYCOSYLATION oxidative stress 

分 类 号：TQ241.2[化学工程—有机化工] TB34[一般工业技术—材料科学与工程]