Structure and dynamics of the CGRP receptor in apo and peptide-bound forms

Tracy M Josephs; Matthew J Belousoff; Yi-Lynn Liang; Sarah J Piper; Jianjun Cao; Daniel J Garama; Katie Leach; Karen J Gregory; Arthur Christopoulos; Debbie L Hay; Radostin Danev; Denise Wootten; Patrick M Sexton

doi:10.1126/science.abf7258

Structure and dynamics of the CGRP receptor in apo and peptide-bound forms

Science. 2021 Apr 9;372(6538):eabf7258. doi: 10.1126/science.abf7258. Epub 2021 Feb 18.

Authors

Affiliations

¹ Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville 3052, Victoria, Australia.
² Hudson Institute of Medical Research, Clayton 3168, Victoria, Australia.
³ Department of Molecular and Translational Science, Monash University, Clayton 3168, Victoria, Australia.
⁴ Department of Pharmacology and Toxicology, University of Otago, Dunedin 9054, New Zealand.
⁵ Graduate School of Medicine, University of Tokyo, N415, 7-3-1 Hongo, Bunkyo-ku, 113-0033 Tokyo, Japan. [email protected] [email protected] [email protected].
⁶ Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville 3052, Victoria, Australia. [email protected] [email protected] [email protected].

^# Contributed equally.

PMID: 33602864
DOI: 10.1126/science.abf7258

Abstract

G protein-coupled receptors (GPCRs) are key regulators of information transmission between cells and organs. Despite this, we have only a limited understanding of the behavior of GPCRs in the apo state and the conformational changes upon agonist binding that lead to G protein recruitment and activation. We expressed and purified unmodified apo and peptide-bound calcitonin gene-related peptide (CGRP) receptors from insect cells to determine their cryo-electron microscopy (cryo-EM) structures, and we complemented these with analysis of protein conformational dynamics using hydrogen-deuterium exchange mass spectrometry and three-dimensional variance analysis of the cryo-EM data. Together with our previously published structure of the active, Gs-bound CGRP receptor complex, our work provides insight into the mechanisms of class B1 GPCR activation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoproteins / chemistry
Apoproteins / metabolism
Calcitonin Gene-Related Peptide / chemistry*
Calcitonin Gene-Related Peptide / metabolism
Calcitonin Receptor-Like Protein / chemistry
Cell Line
Cell Membrane / metabolism
Cryoelectron Microscopy
GTP-Binding Protein alpha Subunits, Gs / chemistry
GTP-Binding Protein alpha Subunits, Gs / metabolism
Humans
Hydrogen Deuterium Exchange-Mass Spectrometry
Ligands
Models, Molecular
Moths
Protein Binding
Protein Conformation
Protein Interaction Domains and Motifs
Protein Structure, Secondary
Receptor Activity-Modifying Protein 1 / chemistry
Receptor Activity-Modifying Protein 1 / metabolism
Receptors, Calcitonin Gene-Related Peptide / chemistry*
Receptors, Calcitonin Gene-Related Peptide / metabolism*

Substances

Apoproteins
CALCRL protein, human
Calcitonin Receptor-Like Protein
Ligands
RAMP1 protein, human
Receptor Activity-Modifying Protein 1
Receptors, Calcitonin Gene-Related Peptide
GTP-Binding Protein alpha Subunits, Gs
Calcitonin Gene-Related Peptide