Cryo-EM structure of the active, Gs-protein complexed, human CGRP receptor

Yi-Lynn Liang; Maryam Khoshouei; Giuseppe Deganutti; Alisa Glukhova; Cassandra Koole; Thomas S Peat; Mazdak Radjainia; Jürgen M Plitzko; Wolfgang Baumeister; Laurence J Miller; Deborah L Hay; Arthur Christopoulos; Christopher A Reynolds; Denise Wootten; Patrick M Sexton

doi:10.1038/s41586-018-0535-y

Cryo-EM structure of the active, G_s-protein complexed, human CGRP receptor

Nature. 2018 Sep;561(7724):492-497. doi: 10.1038/s41586-018-0535-y. Epub 2018 Sep 12.

Authors

Yi-Lynn Liang^#¹, Maryam Khoshouei^#^{2

3}, Giuseppe Deganutti⁴, Alisa Glukhova¹, Cassandra Koole¹, Thomas S Peat⁵, Mazdak Radjainia^{1

6}, Jürgen M Plitzko², Wolfgang Baumeister², Laurence J Miller^{1

7}, Deborah L Hay^{8

9}, Arthur Christopoulos¹, Christopher A Reynolds⁴, Denise Wootten^{10

11}, Patrick M Sexton^{12

13}

Affiliations

¹ Drug Discovery Biology and Department of Pharmacology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
² Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
³ Novartis Institutes for Biomedical Research, Novartis Pharma, Basel, Switzerland.
⁴ School of Biological Sciences, University of Essex, Colchester, UK.
⁵ CSIRO Biomedical Manufacturing, Melbourne, Victoria, Australia.
⁶ Thermo Fisher Scientific, Eindhoven, The Netherlands.
⁷ Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Scottsdale, AZ, USA.
⁸ School of Biological Sciences, University of Auckland, Auckland, New Zealand.
⁹ Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, New Zealand.
¹⁰ Drug Discovery Biology and Department of Pharmacology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia. [email protected].
¹¹ School of Pharmacy, Fudan University, Shanghai, China. [email protected].
¹² Drug Discovery Biology and Department of Pharmacology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia. [email protected].
¹³ School of Pharmacy, Fudan University, Shanghai, China. [email protected].

^# Contributed equally.

Abstract

Calcitonin gene-related peptide (CGRP) is a widely expressed neuropeptide that has a major role in sensory neurotransmission. The CGRP receptor is a heterodimer of the calcitonin receptor-like receptor (CLR) class B G-protein-coupled receptor and a type 1 transmembrane domain protein, receptor activity-modifying protein 1 (RAMP1). Here we report the structure of the human CGRP receptor in complex with CGRP and the G_s-protein heterotrimer at 3.3 Å global resolution, determined by Volta phase-plate cryo-electron microscopy. The receptor activity-modifying protein transmembrane domain sits at the interface between transmembrane domains 3, 4 and 5 of CLR, and stabilizes CLR extracellular loop 2. RAMP1 makes only limited direct contact with CGRP, consistent with its function in allosteric modulation of CLR. Molecular dynamics simulations indicate that RAMP1 provides stability to the receptor complex, particularly in the positioning of the extracellular domain of CLR. This work provides insights into the control of G-protein-coupled receptor function.

MeSH terms

Binding Sites
Calcitonin Gene-Related Peptide / chemistry
Calcitonin Gene-Related Peptide / metabolism*
Calcitonin Receptor-Like Protein / chemistry
Calcitonin Receptor-Like Protein / metabolism
Calcitonin Receptor-Like Protein / ultrastructure*
Cryoelectron Microscopy*
GTP-Binding Protein alpha Subunits, Gs / chemistry
GTP-Binding Protein alpha Subunits, Gs / metabolism*
GTP-Binding Protein alpha Subunits, Gs / ultrastructure*
Humans
Molecular Dynamics Simulation
Protein Domains
Protein Stability
Receptor Activity-Modifying Protein 1 / chemistry
Receptor Activity-Modifying Protein 1 / metabolism
Receptor Activity-Modifying Protein 1 / ultrastructure*
Receptors, Calcitonin Gene-Related Peptide / chemistry
Receptors, Calcitonin Gene-Related Peptide / metabolism*
Receptors, Calcitonin Gene-Related Peptide / ultrastructure*
ras Proteins / chemistry
ras Proteins / metabolism

Substances

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

Grants and funding

BB/M006883/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom