Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics
Sang-min Lee, Yejin Jeong, John Simms, Margaret L. Warner, David R. Poyner, Ka Young Chung, Augen A. Pioszak*
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.
Journal of Molecular Biology
https://doi.org/10.1016/j.jmb.2020.01.028
Published - 27 Mar 2020
© 2020 The Author(s). CC BY
Funding: NIH grant R01GM104251 to AAP, the National Research Foundation of Korea grant NRF-2018R1A2B6001554 to KYC, and use of the crystal imaging microscope was supported by NIH COBRE award P20GM103640. DRP and JS were supported by the BBSRC grant BB/M007529/1.
G protein-coupled receptor (GPCR)
N-linked glycosylation
dynamic allostery
ligand binding kinetics
peptide hormone
10.1016/j.jmb.2020.01.028Licence: CC BY
Calcitonin receptor N-glycosylation
© 2020 The Author(s). Published by Elsevier Ltd.
Accepted author manuscript, 5.99 MBLicence: CC BY
https://linkinghub.elsevier.com/retrieve/pii/S0022283620300929
Fingerprint Dive into the research topics of 'Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics'. Together they form a unique fingerprint.
Calcitonin Receptors Medicine & Life Sciences
Peptide Hormones Medicine & Life Sciences
Glycosylation Medicine & Life Sciences
Polysaccharides Medicine & Life Sciences
Hormones Medicine & Life Sciences
G-Protein-Coupled Receptors Medicine & Life Sciences
Islet Amyloid Polypeptide Receptor Medicine & Life Sciences
Binding Sites Medicine & Life Sciences
Data molecular dynamic simulations of CTR in "Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics"
Simms, J. (Creator) & Poyner, D. (Creator), Aston Data Explorer, 6 Aug 2019
DOI: 10.17036/researchdata.aston.ac.uk.00000433, https://www.sciencedirect.com/science/article/pii/S0022283620300929
Lee, S., Jeong, Y., Simms, J., Warner, M. L., Poyner, D. R., Chung, K. Y., & Pioszak, A. A. (2020). Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics. Journal of Molecular Biology , 432(7), 1996-2014. https://doi.org/10.1016/j.jmb.2020.01.028
Lee, Sang-min ; Jeong, Yejin ; Simms, John ; Warner, Margaret L. ; Poyner, David R. ; Chung, Ka Young ; Pioszak, Augen A. / Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics. In: Journal of Molecular Biology . 2020 ; Vol. 432, No. 7. pp. 1996-2014.
@article{9d71c1a5d3994b89bd908aa1ce9b6a93,
title = "Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics",
abstract = "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 {\AA} 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.",
keywords = "G protein-coupled receptor (GPCR), N-linked glycosylation, dynamic allostery, ligand binding kinetics, peptide hormone",
author = "Sang-min Lee and Yejin Jeong and John Simms and Warner, {Margaret L.} and Poyner, {David R.} and Chung, {Ka Young} and Pioszak, {Augen A.}",
note = "{\textcopyright} 2020 The Author(s). CC BY Funding: NIH grant R01GM104251 to AAP, the National Research Foundation of Korea grant NRF-2018R1A2B6001554 to KYC, and use of the crystal imaging microscope was supported by NIH COBRE award P20GM103640. DRP and JS were supported by the BBSRC grant BB/M007529/1.",
doi = "10.1016/j.jmb.2020.01.028",
journal = "Journal of Molecular Biology ",
Lee, S, Jeong, Y, Simms, J, Warner, ML, Poyner, DR, Chung, KY & Pioszak, AA 2020, 'Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics', Journal of Molecular Biology , vol. 432, no. 7, pp. 1996-2014. https://doi.org/10.1016/j.jmb.2020.01.028
Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics. / Lee, Sang-min; Jeong, Yejin; Simms, John; Warner, Margaret L.; Poyner, David R.; Chung, Ka Young; Pioszak, Augen A.
In: Journal of Molecular Biology , Vol. 432, No. 7, 27.03.2020, p. 1996-2014.
T1 - Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics
AU - Lee, Sang-min
AU - Jeong, Yejin
AU - Simms, John
AU - Warner, Margaret L.
AU - Poyner, David R.
AU - Chung, Ka Young
AU - Pioszak, Augen A.
N1 - © 2020 The Author(s). CC BY Funding: NIH grant R01GM104251 to AAP, the National Research Foundation of Korea grant NRF-2018R1A2B6001554 to KYC, and use of the crystal imaging microscope was supported by NIH COBRE award P20GM103640. DRP and JS were supported by the BBSRC grant BB/M007529/1.
N2 - 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.
AB - 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.
KW - G protein-coupled receptor (GPCR)
KW - N-linked glycosylation
KW - dynamic allostery
KW - ligand binding kinetics
KW - peptide hormone
UR - https://linkinghub.elsevier.com/retrieve/pii/S0022283620300929
U2 - 10.1016/j.jmb.2020.01.028
DO - 10.1016/j.jmb.2020.01.028
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
Lee S, Jeong Y, Simms J, Warner ML, Poyner DR, Chung KY et al. Calcitonin receptor N-glycosylation enhances peptide hormone affinity by controlling receptor dynamics. Journal of Molecular Biology . 2020 Mar 27;432(7):1996-2014. https://doi.org/10.1016/j.jmb.2020.01.028