Xiao Kunhong, Chung Jeannie, Wall Alissa
a Department of Pharmacology and Chemical Biology , University of Pittsburgh , Pittsburgh , PA , USA and.
b Department of Medicine , Duke University Medical Center , Durham , NC , USA.
J Recept Signal Transduct Res. 2015 Jun;35(3):213-9. doi: 10.3109/10799893.2015.1072979. Epub 2015 Oct 12.
Mass spectrometry (MS)-based proteomics is an unrivaled tool for studying complex biological systems and diseases in the post-genomic era. In recent years, MS has emerged as a powerful structural biological tool to characterize protein conformation and conformational dynamics. The advantages of MS in structural studies are most evident for membrane proteins such as GPCRs (G protein-coupled receptors), where other well-established structural methods such as X-ray crystallography and NMR remain challenging. For proteins with available high-resolution structures, MS-based structural strategies can provide valuable, previously inaccessible information on protein conformational changes and dynamics, protein motion/flexibility, ligand-protein binding, and protein-protein interfaces. In the past several years, we have developed and adapted a number of MS-based structural approaches, such as CDSiL-MS (Conformational changes and Dynamics using Stable-isotope Labeling and MS), CXMS (Crosslinking/MS) and HDXMS (Hydrogen-Deuterium Exchange MS), to study protein structures and conformational dynamics in human β2-adrenegic receptor (β2AR) signaling. In this mini-review, we will highlight several examples demonstrating the power of MS in structural analysis to better elucidate the structural basis of GPCR signaling, particularly through the β-arrestin-mediated GPCR signaling pathway.
基于质谱（MS）的蛋白质组学是后基因组时代研究复杂生物系统和疾病的无与伦比的工具。近年来，质谱已成为一种强大的结构生物学工具，用于表征蛋白质构象和构象动力学。质谱在结构研究中的优势在膜蛋白如G蛋白偶联受体（GPCRs）中最为明显，而其他成熟的结构方法如X射线晶体学和核磁共振仍具有挑战性。对于具有高分辨率结构的蛋白质，基于质谱的结构策略可以提供有关蛋白质构象变化和动力学、蛋白质运动/灵活性、配体-蛋白质结合以及蛋白质-蛋白质界面的有价值的、以前无法获得的信息。在过去几年中，我们开发并采用了多种基于质谱的结构方法，如CDSiL-MS（使用稳定同位素标记和质谱的构象变化和动力学）、CXMS（交联/质谱）和HDXMS（氢-氘交换质谱），来研究人β2肾上腺素能受体（β2AR）信号传导中的蛋白质结构和构象动力学。在本综述中，我们将重点介绍几个例子，展示质谱在结构分析中的强大功能，以更好地阐明GPCR信号传导的结构基础，特别是通过β-抑制蛋白介导的GPCR信号通路。
