Endo-M Mediated Chemoenzymatic Approach Enables Reversible Glycopeptide Labeling for O-GlcNAcylation Analysis

被引：19

作者：

Chen, Yao ^{[1
,2
]}

Tang, Feng ^{[2
,3
]}

Qin, Hongqiang ^{[1
,2
]}

Yue, Xuyang ^{[1
]}

Nie, Yongzhan ^{[4
,5
]}

Huang, Wei ^{[2
,3
]}

Ye, Mingliang ^{[1
,2
]}

机构：

[1] Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

[2] Univ Chinese Acad Sci, Beijing 101408, Peoples R China

[3] Chinese Acad Sci, CAS Ctr Excellence Mol Cell Sci, Ctr Biotherapeut Discovery Res, Shanghai Inst Mat Med,CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China

[4] Fourth Mil Med Univ, Natl Clin Res Ctr Digest Dis, State Key Lab Canc Biol, Xian 710032, Peoples R China

[5] Fourth Mil Med Univ, Xijing Hosp Digest Dis, Xian 710032, Peoples R China

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2022年 / 61卷 / 23期

基金：

中国国家自然科学基金;

关键词：

Chemoenzymatic Labeling; Glycopeptide Enrichment; Glycoproteomics; Hydrophilic Interaction Liquid Chromatography; O-GlcNAcylation; BETA-N-ACETYLGLUCOSAMINE; MODIFIED PROTEINS; CROSS-TALK; GLYCOSYLATION; PHOSPHORYLATION; TRANSCRIPTION; DYNAMICS; STRESS;

D O I：

10.1002/anie.202117849

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

To selectively enrich O-linked beta-N-acetylglucosamine (O-GlcNAc) peptides in their original form from complex samples, we report the first reversible chemoenzymatic labeling approach for proteomic analysis. In this strategy, the O-GlcNAc moieties are ligated with long N-glycans using an Endo-M mutant, which enables the enrichment of the labeled glycopeptides by hydrophilic interaction liquid chromatography (HILIC). The attached glycans on the enriched glycopeptides are removed by wild-type Endo-M/S to restore the O-GlcNAc moiety. Compared with classic chemoenzymatic labeling, this approach enables the tag-free identification, and eliminates the interference of bulky tags in glycopeptide detection. This approach presents a unique avenue for the proteome-wide analysis of protein O-GlcNAcylation to promote its mechanism research.

引用

页数：8

共 3 条

[1] Quantitative analysis of O-GlcNAcylation in combination with isobaric tag labeling and chemoenzymatic enrichment
Tsumoto, Hiroki
Akimoto, Yoshihiro
Endo, Tamao
Miura, Yuri
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2017, 27 (22) : 5022 - 5026
[2] Determination of N-glycans in glycoproteins using chemoenzymatic labeling with Endo-M N175Q
Zhang, Si-ran
Yu, Ying-lan
Xu, Chang-shuai
Jin, Dongri
Lee, Yong-Ill
MICROCHEMICAL JOURNAL, 2017, 130 : 390 - 399
[3] An approach to the precise chemoenzymatic synthesis of 13C-labeled sialyloligosaccharide on an intact glycoprotein:: A novel one-pot [3-13C]-labeling method for sialic acid analogues by control of the reversible aldolase reaction, enzymatic synthesis of [3-13C]-NeuAc-α-(2→3)-[U-13C]-Gal-β-(1→4)-GlcNAc-β- sequence onto glycoprotein, and its conformational analysis by developed NMR techniques
Miyazaki, T
Sato, H
Sakakibara, T
Kajihara, Y
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (24) : 5678 - 5694

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