Ultraviolet Photodissociation Permits Comprehensive Characterization of O-Glycopeptides Cleaved with O-Glycoprotease IMPa.
Ultraviolet Photodissociation Permits Comprehensive Characterization of O-Glycopeptides Cleaved with O-Glycoprotease IMPa.
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紫外线光解可以对 O-糖蛋白酶 IMPa 切割的 O-糖肽进行全面表征。
DOI：
10.1021/acs.analchem.3c01111
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发表时间：
2023
影响因子：
7.4
通讯作者：
Brodbelt,JenniferS
中科院分区：
文献类型：
--
作者：
Helms,Amanda;Escobar,EdwinE;Vainauskas,Saulius;Taron,ChristopherH;Brodbelt,JenniferS
完整的O-糖基表征，包括肽的鉴定、糖基的定位和聚糖的作图，由于O-聚糖分析的技术挑战，一直是O-糖蛋白质组学中的一个持续挑战。多糖基化肽由于其潜在的异质性而构成更大的挑战。紫外光解离（UVPD）可以定位多种翻译后修饰，非常适合于聚糖的表征。基于结合使用O-糖蛋白酶IMPa和HCD触发的UVPD的策略，对三种糖蛋白进行了评估，以完整表征O-糖肽。该方法定位了单个糖肽上的多个相邻或近端O-糖位点，并在S218处鉴定了依那西普上先前未知的糖位点。9种不同的糖型被表征为来自依那西普的多糖基化肽。UVPD的性能进行了比较的HCD和EThcD的O-糖位点的定位和组成肽和聚糖的表征。
CompleteO-glycosite characterization, including identification of the peptides, localization of the glycosites, and mapping of the glycans, has been a persistent challenge inO-glycoproteomics owing to the technical challenges surroundingO-glycan analysis. Multi-glycosylated peptides pose an even greater challenge owing to their potential heterogeneity. Ultraviolet photodissociation (UVPD) can localize multiple post-translational modifications and is well-suited for the characterization of glycans. Three glycoproteins were assessed based on a strategy combining the use ofO-glycoprotease IMPa and HCD-triggered UVPD for the complete characterization ofO-glycopeptides. This approach localized multiple adjacent or proximalO-glycosites on individual glycopeptides and identified a previously unknown glycosite on etanercept at S218. Nine different glycoforms were characterized as a multi-glycosylated peptide from etanercept. The performance of UVPD was compared to that of HCD and EThcD for the localization ofO-glycosites and the characterization of the constituent peptides and glycans.