揭示粘蛋白 O-糖的多样性：从 SmE 粘蛋白酶和紫外光解离质谱法中获得的启示

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2024-11-22 DOI:10.1021/acs.analchem.4c02011

Amanda Helms, Vincent Chang, Stacy A. Malaker, Jennifer S. Brodbelt

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引用次数: 0

摘要

粘蛋白结构域蛋白是与癌症和其他疾病密切相关的糖蛋白，解读粘蛋白结构域蛋白的 O 型糖基化模式和丰度仍是一项持续的挑战。糖基化的宏观和微观异质性使分析变得更加复杂，这促使人们开发新的策略来确定这类糖蛋白的结构特征。在这里，我们将使用一种靶向蛋白酶--来自马氏沙雷氏菌的 Enhancin（SmE）--消化粘蛋白结构域蛋白与紫外光解离（UVPD）质谱法结合起来，以推进糖图谱绘制并阐明密集糖基化粘蛋白的 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-糖基化形式。此外，UVPD还阐明了MUC-16的几种糖基化形式，并有助于发现MUC-1串联重复序列的O-糖基化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Unraveling O-Glycan Diversity of Mucins: Insights from SmE Mucinase and Ultraviolet Photodissociation Mass Spectrometry

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Unraveling O-Glycan Diversity of Mucins: Insights from SmE Mucinase and Ultraviolet Photodissociation Mass Spectrometry

Deciphering the pattern and abundance of O-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 Serratia marcescens (SmE), with ultraviolet photodissociation (UVPD) mass spectrometry to advance glycan mapping and elucidation of O-glycosylation trends of densely glycosylated mucin proteins. UVPD facilitates identification of O-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 O-glycosylation across tandem repeats of MUC-1.

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.