Ligand Binding to a G Protein-coupled Receptor Captured in a Mass Spectrometer

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2017 Jun 21

PMID 28630934

Citations 29

Authors

Hsin-Yung Yen

Jonathan T S Hopper

Idlir Liko

Timothy M Allison

Ya Zhu

Dejian Wang

Monika Stegmann

Shabaz Mohammed

Beili Wu

Carol V Robinson

Affiliations

Soon will be listed here.

Abstract

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 P2Y. 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.

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