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.

doi:10.1016/j.jmb.2020.01.028

Title: Calcitonin Receptor N-Glycosylation Enhances Peptide Hormone Affinity by Controlling Receptor Dynamics

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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 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.

Authors:

Lee, Sang-Min ^[1]; Jeong, Yejin ^[2]; Simms, John ^[3]; Warner, Margaret L. ^[4]; Poyner, David R. ^[3]; Chung, Ka Young ^[3]; Pioszak, Augen A. ^[4]

Oklahoma City Univ. Health Sciences Center, Oklahoma City, OK (United States); High Point Univ., High Point, NC (United States)
Sungkyunkwan Univ., Suwon (Korea)
Aston Univ., Aston Triangle, Birmingham (United Kingdom)
Oklahoma City Univ. Health Sciences Center, Oklahoma City, OK (United States)

Publication Date:: 2020-03-27

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Org.:: USDOE Office of Science (SC); National Research Foundation of Korea (NRF); National Institutes of Health (NIH); Michigan Economic Development Corporation

OSTI Identifier:: 1657570

Grant/Contract Number:: AC02-06CH11357; 085P1000817; R01GM104251; NRF-2018R1A2B6001554; BB/M007529/1

Resource Type:: Accepted Manuscript

Journal Name:: Journal of Molecular Biology

Additional Journal Information:: Journal Volume: 432; Journal Issue: 7; Journal ID: ISSN 0022-2836

Publisher:: Elsevier

Country of Publication:: United States

Language:: ENGLISH

Subject:: 59 BASIC BIOLOGICAL SCIENCES; G protein-coupled receptor (GPCR); N-linked glycosylation; peptide hormone; ligand binding kinetics; dynamic allostery

Citation Formats


                    Lee, Sang-Min, Jeong, Yejin, Simms, John, Warner, Margaret L., Poyner, David R., Chung, Ka Young, and Pioszak, Augen A. Calcitonin Receptor N-Glycosylation Enhances Peptide Hormone Affinity by Controlling Receptor Dynamics.  United States: N. p., 2020. 
Web.  doi:10.1016/j.jmb.2020.01.028.

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                    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.  United States.  https://doi.org/10.1016/j.jmb.2020.01.028

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                    Lee, Sang-Min, Jeong, Yejin, Simms, John, Warner, Margaret L., Poyner, David R., Chung, Ka Young, and Pioszak, Augen A. Fri .  
"Calcitonin Receptor N-Glycosylation Enhances Peptide Hormone Affinity by Controlling Receptor Dynamics".  United States.  https://doi.org/10.1016/j.jmb.2020.01.028.  https://www.osti.gov/servlets/purl/1657570.

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@article{osti_1657570,

  title        = {Calcitonin Receptor N-Glycosylation Enhances Peptide Hormone Affinity by Controlling Receptor Dynamics},

  author       = {Lee, Sang-Min and Jeong, Yejin and Simms, John and Warner, Margaret L. and Poyner, David R. and Chung, Ka Young and Pioszak, Augen A.},

  abstractNote = {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.},

  doi          = {10.1016/j.jmb.2020.01.028},

  journal      = {Journal of Molecular Biology},

  number       = 7,

  volume       = 432,

  place        = {United States},

  year         = {2020},

  month        = {3}

}

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