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.

中文翻译：

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用于蛋白质和复合物高阶结构表征的天然自上而下质谱法

结构生物学研究的进展导致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的高需求。这种需求显着增加了人们对原生质谱 (nMS)，特别是过去十年中原生自上而下质谱 (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 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、串联碎片技术以及用于 nTDMS 的质量分析仪和软件/分析管道。我们重点介绍 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、大分子等领域的广泛应用示例。机器（核糖体、核小体、蛋白体和病毒）到内源性蛋白质复合物。讨论了用于分析重组和生物样品中蛋白质和蛋白质组装体的 nTDMS 方法的挑战、潜力和前景。