title: The power of mass spectrometry in structural characterization of GPCR signaling.
abstract: 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.
authors.affiliation: 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.
authors.forename: Kunhong
authors.initials: K
authors.lastname: Xiao
authors.affiliation: b Department of Medicine , Duke University Medical Center , Durham , NC , USA.
authors.forename: Jeannie
authors.initials: J
authors.lastname: Chung
authors.affiliation: b Department of Medicine , Duke University Medical Center , Durham , NC , USA.
authors.forename: Alissa
authors.initials: A
authors.lastname: Wall
mesh_terms: D000818:Animals; D003461:Crystallography / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008954:Models, Biological; D008956:Models, Chemical; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*
chemical_list: D043562:Receptors, G-Protein-Coupled
keywords: G protein-coupled receptor; GPCR; HDXMS; arrestin; mass spectrometry; protein structure; proteomics; signaling