解析粘蛋白的聚糖多样性：来自SmE粘蛋白酶和紫外光解离质谱的见解
Unraveling -Glycan Diversity of Mucins: Insights from SmE Mucinase and Ultraviolet Photodissociation Mass Spectrometry.
作者信息
Helms Amanda, Chang Vincent, Malaker Stacy A, Brodbelt Jennifer S
机构信息
Department of Chemistry, University of Texas, Austin, Texas 78712, United States.
Department of Chemistry, Yale University, New Haven, Connecticut 06511, United States.
出版信息
Anal Chem. 2024 Dec 10;96(49):19230-19237. doi: 10.1021/acs.analchem.4c02011. Epub 2024 Nov 22.
Deciphering the pattern and abundance of -glycosylation of mucin domain proteins, glycoproteins heavily implicated in cancer and other diseases, remains an ongoing challenge. Both the macro- and microheterogeneity of glycosylation complicates the analysis, motivating the development of new strategies for structural characterization of this diverse class of glycoproteins. Here we combine digestion of mucin domain proteins using a targeted protease, Enhancin from (SmE), with ultraviolet photodissociation (UVPD) mass spectrometry to advance glycan mapping and elucidation of -glycosylation trends of densely glycosylated mucin proteins. UVPD facilitates identification of -glycoforms of mucin domain proteins TIM-1, MUC-1 and MUC-16. Additionally, UVPD elucidates several glycoforms of MUC-16 and contributes to the discovery of -glycosylation across tandem repeats of MUC-1.
解读粘蛋白结构域蛋白的O-糖基化模式和丰度仍是一项持续的挑战，这类糖蛋白与癌症和其他疾病密切相关。糖基化的宏观和微观异质性使分析变得复杂，这推动了针对这类多样糖蛋白进行结构表征的新策略的开发。在此，我们将使用靶向蛋白酶（来自斯氏嗜银菌的增强蛋白，SmE）对粘蛋白结构域蛋白进行酶解，与紫外光解离（UVPD）质谱相结合，以推进聚糖图谱分析，并阐明高度糖基化的粘蛋白蛋白的O-糖基化趋势。UVPD有助于鉴定粘蛋白结构域蛋白TIM-1、MUC-1和MUC-16的O-糖型。此外，UVPD还阐明了MUC-16的几种糖型，并有助于发现MUC-1串联重复序列中的O-糖基化。
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