蛋白质糖基化修饰的非变性构象分辨质谱研究进展  

Recent progress of protein glycosylation characterization utilizing native conformer-resolved mass spectrometry

作　　者：贾翼菲 王雅梅 李功玉 JIA Yifei;WANG Yamei;LI Gongyu(Tianjin Key Laboratory of Biosensing and Molecular Recognition,Research Center for Analytical Science,College of Chemistry,Nankai University,Tianjin 300071;Haihe Laboratory of Sustainable Chemical Transformations,Tianjin 300192,China)

机构地区：[1]南开大学化学学院,分析科学研究中心,天津市生物传感与分子识别重点实验室,天津300071 [2]物质绿色创造与制造海河实验室,天津300192

出　　处：《中国药科大学学报》2023年第6期674-681,共8页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.22104064,No.22293030,No.22293032);国家重点研发计划青年科学家项目资助(No.2022YFA1305200)。

摘　　要：糖基化是蛋白质最重要的翻译后修饰之一,能够调控蛋白质的电荷态、结构及分子间相互作用等,进而影响其功能。糖基化的高度异质性导致传统的结构解析方法很难对糖蛋白进行全面表征。随着分析技术的发展,质谱在糖蛋白结构解析中发挥了重要作用。蛋白质组学质谱技术可在多肽水平上对复杂、低丰度蛋白质糖基化修饰的化学组成与位点信息进行鉴定。非变性质谱技术(native mass spectrometry,nMS)则直接在完整蛋白水平上揭示聚糖异质性及其对蛋白高级结构和相互作用的调控效应。作为结构质谱的代表性技术,基于离子淌度的nMS受益于离子淌度仪的构象分辨能力和构象去折叠功能,能够在非变性质谱的基础上提供离子的三维动态结构信息,为异构体结构快速鉴定提供不可替代的解决方案。本文重点介绍了两种新兴离子淌度质谱技术,即非变性动态构象分辨质谱技术和糖型分辨结构质谱技术,并以3种常见蛋白体系为例,介绍其在糖蛋白构象研究领域的最新进展。Glycosylation of proteins,one of the most prevalent and complex post-translational modifications occurring in nature,plays a crucial role in regulating protein net charge,conformation,binding properties and,ultimately,biological function.Traditional structural techniques are not amenable for glycoproteins due to the inherent heterogeneity of oligosaccharides.With the advances in analytical technique,mass spectrometry displays an increasingly crucial role in elucidating the structure of glycoproteins.Mass spectrometry-based proteomic technique can dissect the chemical composition and site information of low-abundance glycosylation at the peptide level.Instead,native mass spectrometry(nMS)can analyze intact glycoproteins while maintaining the information for glycan heterogeneity,and the insights into the regulatory effects of glycosylation on protein higher order structures and interactions with other proteins or ligands.As a representative structural mass spectrometry tool,ion mobility-based nMS strategy is powered by its conformer-resolving capability and by the feasibility of conformer manipulation through collision-induced unfolding.Consequently,native IM-MS analysis can provide rich information of dynamic protein conformations,allowing for the rapid identification and differ-entiation of protein isoforms in an unprecedented manner.In this review,we briefly introduced two emerging native IM-MS analytical modes,dynamic conformer-resolving mode and glycoform-resolving mode.Besides,we also discussed the recent progress of conformational and topological characterization of intact glycoproteins with three typical model systems based on two above-mentioned emerging modes of native IM-MS.

关 键 词：蛋白糖基化 非变性质谱技术 构象分辨质谱 离子淌度质谱 碰撞诱导去折叠 

分 类 号：R363[医药卫生—病理学]