Liu Ruijie, Xia Shujun, Li Huilin
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China.
Guangdong Key Laboratory of Chiral Molecule and Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China.
Mass Spectrom Rev. 2023 Sep-Oct;42(5):1876-1926. doi: 10.1002/mas.21793. Epub 2022 Jun 27.
Progress in structural biology research has led to a high demand for powerful and yet complementary analytical tools for structural characterization of proteins and protein complexes. This demand has significantly increased interest in native mass spectrometry (nMS), particularly native top-down mass spectrometry (nTDMS) in the past decade. This review highlights recent advances in nTDMS for structural research of biological assemblies, with a particular focus on the extra multi-layers of information enabled by TDMS. We include a short introduction of sample preparation and ionization to nMS, tandem fragmentation techniques as well as mass analyzers and software/analysis pipelines used for nTDMS. We highlight unique structural information offered by nTDMS and examples of its broad range of applications in proteins, protein-ligand interactions (metal, cofactor/drug, DNA/RNA, and protein), therapeutic antibodies and antigen-antibody complexes, membrane proteins, macromolecular machineries (ribosome, nucleosome, proteosome, and viruses), to endogenous protein complexes. The challenges, potential, along with perspectives of nTDMS methods for the analysis of proteins and protein assemblies in recombinant and biological samples are discussed.
结构生物学研究的进展使得对用于蛋白质和蛋白质复合物结构表征的强大且互补的分析工具产生了很高的需求。在过去十年中，这种需求显著提高了人们对天然质谱（nMS），特别是天然自上而下质谱（nTDMS）的兴趣。本综述重点介绍了nTDMS在生物组装体结构研究方面的最新进展，特别关注TDMS所提供的额外多层信息。我们简要介绍了nMS的样品制备和电离、串联碎裂技术以及用于nTDMS的质量分析器和软件/分析流程。我们强调了nTDMS提供的独特结构信息及其在蛋白质、蛋白质-配体相互作用（金属、辅因子/药物、DNA/RNA和蛋白质）、治疗性抗体和抗原-抗体复合物、膜蛋白、大分子机器（核糖体、核小体、蛋白酶体和病毒）以及内源性蛋白质复合物等广泛应用中的实例。讨论了nTDMS方法在重组和生物样品中分析蛋白质和蛋白质组装体时面临的挑战、潜力以及前景。
