Ligand binding to a G protein–coupled receptor captured in a mass spectrometer (2017)
AuthorsYen HY, Hopper JTS, Liko I, Allison TM, Zhu Y, Wang D, Stegmann M, Mohammed S, Wu B, Robinson CVshow all
Copyright © 2017 The Authors, some rights reserved. G protein (heterotrimeric guanine nucleotide–binding protein)–coupled receptors belong to the largest family of membrane-embedded cell surface proteins and are involved in a diverse array of physiological processes. Despite progress in the mass spectrometry of membrane protein complexes, G protein–coupled receptors have remained intractable because of their low yield and instability after extraction from cell membranes. We established conditions in the mass spectrometer that preserve noncovalent ligand binding to the human purinergic receptor P2Y1. Results established differing affinities for nucleotides and the drug MRS2500 and link antagonist binding with the absence of receptor phosphorylation. Overall, therefore, our results are consistent with drug binding, preventing the conformational changes that facilitate downstream signaling. More generally, we highlight opportunities for mass spectrometry to probe effects of ligand binding on G protein–coupled receptors.
KeywordsG-protein coupled receptors; ligand binding; mass spectrometry
ANZSRC Fields of Research03 - Chemical Sciences::0304 - Medicinal and Biomolecular Chemistry::030406 - Proteins and Peptides
06 - Biological Sciences::0601 - Biochemistry and Cell Biology::060109 - Proteomics and Intermolecular Interactions (excl. Medical Proteomics)
03 - Chemical Sciences::0306 - Physical Chemistry (incl. Structural)::030606 - Structural Chemistry and Spectroscopy