非变性质谱在非共价蛋白质复合物和结构生物学研究领域的应用.docVIP

非变性质谱在非共价蛋白质复合物和结 构生物学研究领域的应用 武国华 江苏科技大学生物技术学院，农业部蚕桑产品及食用 昆虫质量安全风险评估实验室 摘要： 质谱(MS)是表征蛋白质和蛋白质翻译后修饰(PTM)的一种强大的技术手段， 在现代牛物学研究领域中发挥着越来越重要的作用。近年来，非变性质谱也已成 功用于研究蛋白质非共价相互作用，在研究化学计量、结构功能和蛋白质组装动 力学方面彰显出灵敏、快速、专一及信息量大等优势。这种在结构生物学前沿领 域应用的技术工具发展于20世纪90年代初期，虽然其早期研究主要集中在利用 质谱的质量测定来获得蛋白质复合物中的化学计量等信息，但随着质谱技术和 方法的进一步发展，现在己能够用于揭示蛋白质结构、亚基间连接性和蛋白质动 力学等生命科学问题。本文及后续文章中将对应用非变性质谱研究非共价蛋白质 复合物的概况、最适宜的离子源和质量分析器、实验中可能遇到的问题及应对策 略等进行综述与讨论，并辅以相关的应用实例进行较为详细的介绍。 关键词： 非变性质谱;非共价相互作用;蛋白质复合物;结构生物学; 作者简介：武国华(1963—),男，研究员，江苏省特聘教授。 E-mail:georgew51l@hotmai1.co.uk 收稿日期：2017-07-24 基金：江苏省特聘教授专项基金项目(No.苏教师[2015]17) Applications of Native Mass Spectrometry in Non-covalent Protein Complexes and Struetural Biology Wu Guohua College of Biotechnology, Jiangsu University of Science and Technology, Laboratory of Quality Safety Risk Assessment for Sericultural Products and Edible Insects, Ministry of Agriculture; Abstract： Mass spectrometry ( MS) is currently playing an increasingly important role in biological research as a powerful technique to characterize proteins and protein post~transleitioneil modifications ( PTMs) . In recent years, native or non-dcnaturing mass spectromctry has been successfully used to study non-covalent interactions between protein and other molecules, making it a sensitive, quick, specific and informative technique in studies of stoichiometry, structure-function, and protein assembly dynamics. This physiochemical tool used in the frontiers of structural biology stems from experiments performed in the early 1990 s. Although its initial studies focused on the elucidation of stoichiometry by means of mass determination, developments in MS technology and methodology in the past decades have allowed researchers to address questions such as protein shape, inter~subunit association and protein dynamics. In this emd following articlcs, basic facts, the most suitable ion sources and mass analyzers, possible problems confronted in experiments and coping strategies in using native mass spectrometry