非变性质谱相关技术的研究进展  

Recent Advances of Native Mass Spectrometry Techniques

作　　者：谭聪睿 徐伟[1] TAN Cong-rui;XU Wei(School of Life Science,Beijing Institute of Technology,Beijing 100081,China)

机构地区：[1]北京理工大学生命学院,北京100081

出　　处：《质谱学报》2022年第6期754-767,I0005,共15页Journal of Chinese Mass Spectrometry Society

基　　金：科技部仪器专项(2020YFF01014502);国家自然科学基金(21922401,21827810,61635003)。

摘　　要：蛋白质与其他分子的相互作用几乎在所有的生命活动中起着核心调控作用,这些相互作用力和形成的蛋白质复合物是现代生命科学的研究重点。由于传统的生物物理技术对蛋白质复合物和相互作用的研究存在样品纯度要求高的限制,因此迫切需要新技术的出现,为结构生物学和相互作用组学的研究提供补充。质谱技术可以从原理上对混合样品进行检测,降低对样品纯度的要求,其中非变性质谱展现出强大的连接与互补作用。本文从样品制备、离子源、质量分析器、质谱联用技术等4方面介绍非变性质谱相关技术及近年来的研究进展,并总结分析未来面临的挑战以及发展方向。Nearly all the life activities are regulated by the interactions of the protein with other biomolecules,including protein-protein interactions(PPIs)and protein-ligand interactions(PLIs),therefore understanding interaction and structural characterization of formed complexes represent a key task for modern bioscience.Conventional biophysical techniques like circular dichroism spectroscopy(CD),fluorescence resonance energy transfer(FRET),surface plasmon resonance(SPR),and isothermal titration calorimetry(ITC),are limited in studying protein complexes structures and interactions,considering sample purity is a major issue for obtaining reliable data with conventional methods mentioned above.Additional technologies are required to complement structural biology and interactomics research,while mass spectrometry(MS)can deal with impurities or even mixtures in principle,therefore native mass spectrometry(nMS)is demonstrated as a powerful tool to bridge the gap between structural biology and interactomics research.As the most widespread soft ionization technique,electrospray ionization is utilized in native mass spectrometry,which employs volatile aqueous solution,near physiological pH and gentle desolvation conditions to make the transfer of weakly bound noncovalent complexes into the gas phase while preserving their integrity and native structure possible.In this way,stoichiometry,binding partners,thermodynamics,structural dynamics,and topology information on transferred noncovalent complexes can be acquired.A recent breakthrough in native MS sample preparation demonstrated direct analysis of overexpressed proteins from crude cell lysates,which significantly simplified sample purification procedures.Innovations in mass spectrometry instruments,such as extended m/z range quadrupole stages or modification of Orbitrap instruments allow access to high mass with little compromise in resolving power.This has promoted the native analysis of protein complexes and diverse protein interaction networks to elucidate their architectu

关 键 词：非变性质谱(native mass spectrometry) 电喷雾电离 离子淌度 蛋白质复合物 蛋白质相互作用 

分 类 号：O657.63[理学—分析化学]