CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 116023, Dalian, China.
University of Chinese Academy of Sciences, 101408, Beijing, China.
Angew Chem Int Ed Engl. 2022 Jun 7;61(23):e202117849. doi: 10.1002/anie.202117849. Epub 2022 Mar 30.
To selectively enrich O-linked β-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.
为了从复杂样品中以原始形式选择性地富集 O-连接β-N-乙酰氨基葡萄糖（O-GlcNAc）肽，我们报道了第一个用于蛋白质组分析的可还原的化学酶标记方法。在该策略中，O-GlcNAc 部分通过使用 Endo-M 突变体与长 N-聚糖连接，这使得标记的糖肽能够通过亲水相互作用色谱（HILIC）进行富集。用野生型 Endo-M/S 去除富集的糖肽上的附着聚糖，以恢复 O-GlcNAc 部分。与经典的化学酶标记相比，该方法能够实现无标签的鉴定，并消除了在糖肽检测中大体积标签的干扰。该方法为蛋白质 O-GlcNAcylation 的全蛋白质组分析提供了独特的途径，以促进其机制研究。
