Calcitonin Receptor N-Glycosylation Enhances Peptide Hormone Affinity by Controlling Receptor Dynamics.
G protein-coupled receptor (GPCR)
N-linked glycosylation
dynamic allostery
ligand binding kinetics
peptide hormone
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
27 03 2020
27 03 2020
Historique:
received:
24
08
2019
revised:
27
11
2019
accepted:
27
01
2020
pubmed:
10
2
2020
medline:
28
8
2020
entrez:
10
2
2020
Statut:
ppublish
Résumé
The class B G protein-coupled receptor (GPCR) calcitonin receptor (CTR) is a drug target for osteoporosis and diabetes. N-glycosylation of asparagine 130 in its extracellular domain (ECD) enhances calcitonin hormone affinity with the proximal GlcNAc residue mediating this effect through an unknown mechanism. Here, we present two crystal structures of salmon calcitonin-bound, GlcNAc-bearing CTR ECD at 1.78 and 2.85 Å resolutions and analyze the mechanism of the glycan effect. The N130 GlcNAc does not contact the hormone. Surprisingly, the structures are nearly identical to a structure of hormone-bound, N-glycan-free ECD, which suggested that the GlcNAc might affect CTR dynamics not observed in the static crystallographic snapshots. Hydrogen-deuterium exchange mass spectrometry and molecular dynamics simulations revealed that glycosylation stabilized a β-sheet adjacent to the N130 GlcNAc and the N-terminal α-helix near the peptide-binding site while increasing flexibility of the peptide-binding site turret loop. These changes due to N-glycosylation increased the ligand on-rate and decreased its off-rate. The glycan effect extended to RAMP-CTR amylin receptor complexes and was also conserved in the related CGRP receptor. These results reveal that N-glycosylation can modulate GPCR function by altering receptor dynamics.
Identifiants
pubmed: 32035902
pii: S0022-2836(20)30092-9
doi: 10.1016/j.jmb.2020.01.028
pmc: PMC7225057
mid: NIHMS1557427
pii:
doi:
Substances chimiques
Ligands
0
Receptors, Calcitonin
0
Receptors, G-Protein-Coupled
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1996-2014Subventions
Organisme : NIGMS NIH HHS
ID : P20 GM103640
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA225520
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM104251
Pays : United States
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/M007529/1
Pays : United Kingdom
Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.
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