非变性结构质谱在蛋白提取、离子源开发与构象解析方面的研究进展

非变性条件下的蛋白质研究可以提供更准确的高级结构信息，为“序列-结构-功能”关系解析提供更接近生理条件的分子基础。非变性质谱（native MS）是在非变性条件下对目标蛋白质进行质谱分析的技术。近年来，非变性质谱在蛋白构象、蛋白质非共价相互作用以及蛋白质组装复合物等高阶结构解析中发挥着不可替代的作用，并逐渐成为结构生物学研究的重要支撑方法之一。本文总结了非变性结构质谱在蛋白提取、离子源开发与构象解析方面的发展现状，并提出了非变性结构质谱领域现阶段的技术瓶颈与潜在的解决策略，主要包含原位分析能力和结构分辨率的提升这两方面。

Recent Progress of Native Mass Spectrometry on Protein Extraction,Ion Source Development and Conformational Interpretation

Protein characterization highly depends on its operating conditions, where native or often referred as non-denaturing options will greatly facilitate the collection of more biologically meaningful information. Native mass spectrometry (MS) is such a technique that enables protein analysis under non-denaturing conditions with an affordable time and sample cost. The indispensable roles of native MS-based strategies in the deep dissection of protein conformations, non-covalent interactions, and self-assembly stoichiometry have been seen in past decade, which suggest that native MS has increasingly grown to be a ground technique for the structural biology realm. In this review, we firstly introduced the current method development for protein extraction compatible with native MS experiments, ranging from traditional methods to in situ extraction means, which is key to achieve biologically meaningful native MS results. Then, we focused on the most recent advancements in the native MS-compatible ion source and novel regimes for protein conformational interpretation. Specifically, except the most frequently used nanoelectrospray ion source for native MS, we also listed a variety of emerging ionization regimes including nano-desorption electrospray ionization, submicron emitter tip electrospray, induced nanoelectrospray ionization, step-voltage nanoelectrospray ionization. For protein structural analysis, native MS relies on both charge state distribution and conformational interrogation tool, e.g. ion mobility spectrometry. The structural information can be further enriched when coupled to collision-induced unfolding technique, which was also discussed with most recent developments in data acquisition and visualization methods. Current challenges and potential solutions facing native MS realm in terms of both sampling methods and structural resolution were tentatively discussed.