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

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

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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 500 μg sample.Surprisingly,it allows simultaneous identification of N-glycosites,O-GlcNAc sites,O-GaINAc 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.

glycoproteomicslarge-scale enrichmentO-GlcNAcylationN-glycosylationoxidative stress

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

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Department of Chemistry and Liver Cancer Institute,Zhongshan Hospital,Fudan University,Shanghai 200032,China

Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research,Fudan University,Shanghai 200032,China

2024

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ISSN:
年,卷(期):2024.11(11)