Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, California 90095, USA.
Department of Chemistry and Biochemistry, UCLA/DOE Institute of Genomics and Proteomics, and UCLA Molecular Biology Institute, University of California, Los Angeles, California 90095, USA.
Nat Chem. 2018 Feb;10(2):139-148. doi: 10.1038/nchem.2908. Epub 2018 Jan 1.
Mass spectrometry (MS) has become a crucial technique for the analysis of protein complexes. Native MS has traditionally examined protein subunit arrangements, while proteomics MS has focused on sequence identification. These two techniques are usually performed separately without taking advantage of the synergies between them. Here we describe the development of an integrated native MS and top-down proteomics method using Fourier-transform ion cyclotron resonance (FTICR) to analyse macromolecular protein complexes in a single experiment. We address previous concerns of employing FTICR MS to measure large macromolecular complexes by demonstrating the detection of complexes up to 1.8 MDa, and we demonstrate the efficacy of this technique for direct acquirement of sequence to higher-order structural information with several large complexes. We then summarize the unique functionalities of different activation/dissociation techniques. The platform expands the ability of MS to integrate proteomics and structural biology to provide insights into protein structure, function and regulation.
质谱（MS）已成为分析蛋白质复合物的关键技术。传统的 MS 分析主要研究蛋白质亚基的排列，而蛋白质组学 MS 则侧重于序列鉴定。这两种技术通常是分开进行的，没有利用它们之间的协同作用。在这里，我们描述了一种集成的 native MS 和自上而下的蛋白质组学方法的开发，该方法使用傅里叶变换离子回旋共振（FTICR）在单个实验中分析大分子蛋白质复合物。我们通过证明可以检测高达 1.8 MDa 的复合物来解决之前使用 FTICR MS 测量大分子复合物的问题，并通过几个大复合物展示了该技术直接获取序列到更高阶结构信息的有效性。然后，我们总结了不同活化/解离技术的独特功能。该平台扩展了 MS 将蛋白质组学和结构生物学结合起来的能力，以提供对蛋白质结构、功能和调控的深入了解。
