Molecular insights into peptide agonist engagement with the PTH receptor

Brian P. Cary; Elliot J. Gerrard; Matthew J. Belousoff; Madeleine M. Fletcher; Yan Jiang; Isabella C. Russell; Sarah J. Piper; Denise Wootten; Patrick M. Sexton

doi:10.1016/j.str.2023.04.002

Molecular insights into peptide agonist engagement with the PTH receptor

Brian P. Cary
, Elliot J. Gerrard
, Matthew J. Belousoff
, Madeleine M. Fletcher
, Yan Jiang
, Isabella C. Russell
, Sarah J. Piper
, Denise Wootten
, Patrick M. Sexton

Drug Discovery Biology

Research output: Contribution to journal › Article › Research › peer-review

15 Citations (Scopus)

Abstract

The parathyroid hormone (PTH) 1 receptor (PTH1R) is a G protein-coupled receptor (GPCR) that regulates skeletal development and calcium homeostasis. Here, we describe cryo-EM structures of the PTH1R in complex with fragments of the two hormones, PTH and PTH-related protein, the drug abaloparatide, as well as the engineered tool compounds, long-acting PTH (LA-PTH) and the truncated peptide, M-PTH(1–14). We found that the critical N terminus of each agonist engages the transmembrane bundle in a topologically similar fashion, reflecting similarities in measures of Gαs activation. The full-length peptides induce subtly different extracellular domain (ECD) orientations relative to the transmembrane domain. In the structure bound to M-PTH, the ECD is unresolved, demonstrating that the ECD is highly dynamic when unconstrained by a peptide. High resolutions enabled identification of water molecules near peptide and G protein binding sites. Our results illuminate the action of orthosteric agonists of the PTH1R.

Original language	English
Pages (from-to)	668-676.e5
Number of pages	14
Journal	Structure
Volume	31
Issue number	6
DOIs	https://doi.org/10.1016/j.str.2023.04.002
Publication status	Published - 1 Jun 2023

Keywords

Cryo-EM
g protein-coupled receptor
GPCR
hormone
membrane protein
parathyroid
peptides
PTH

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10.1016/j.str.2023.04.002

Cite this

@article{e4ac50276ce34ee9a65d3e3ad0b242d0,

title = "Molecular insights into peptide agonist engagement with the PTH receptor",

abstract = "The parathyroid hormone (PTH) 1 receptor (PTH1R) is a G protein-coupled receptor (GPCR) that regulates skeletal development and calcium homeostasis. Here, we describe cryo-EM structures of the PTH1R in complex with fragments of the two hormones, PTH and PTH-related protein, the drug abaloparatide, as well as the engineered tool compounds, long-acting PTH (LA-PTH) and the truncated peptide, M-PTH(1–14). We found that the critical N terminus of each agonist engages the transmembrane bundle in a topologically similar fashion, reflecting similarities in measures of Gαs activation. The full-length peptides induce subtly different extracellular domain (ECD) orientations relative to the transmembrane domain. In the structure bound to M-PTH, the ECD is unresolved, demonstrating that the ECD is highly dynamic when unconstrained by a peptide. High resolutions enabled identification of water molecules near peptide and G protein binding sites. Our results illuminate the action of orthosteric agonists of the PTH1R.",

keywords = "Cryo-EM, g protein-coupled receptor, GPCR, hormone, membrane protein, parathyroid, peptides, PTH",

author = "Cary, \{Brian P.\} and Gerrard, \{Elliot J.\} and Belousoff, \{Matthew J.\} and Fletcher, \{Madeleine M.\} and Yan Jiang and Russell, \{Isabella C.\} and Piper, \{Sarah J.\} and Denise Wootten and Sexton, \{Patrick M.\}",

note = "Funding Information: P.M.S. is a Senior Principal Research Fellow (no. 1154434) and D.W. a Senior Research Fellow (no. 1155302) of the National Health and Medical Research Council of Australia. P.M.S. is the Director and D.W. the Monash University Node leader of the Australian Research Council Industrial Transformation Training Centre for Cryo-electron Microscopy of Membrane Proteins (no. IC200100052). This work was funded in part by an NHMRC Program Grant (no. 1150083 ) to P.M.S. High-performance computing was supported by Monash MASSIVE. Cryo-EM samples were imaged at the Bio21 Advanced Microscopy Facility (The University of Melbourne) and at the Monash Ramaciotti Centre for Cryo-Electron Microscopy. The authors thank Dr. Hari Venugopal for assistance with 300 kV data collection at the Monash Ramaciotti Centre. Funding Information: P.M.S. is a Senior Principal Research Fellow (no. 1154434) and D.W. a Senior Research Fellow (no. 1155302) of the National Health and Medical Research Council of Australia. P.M.S. is the Director and D.W. the Monash University Node leader of the Australian Research Council Industrial Transformation Training Centre for Cryo-electron Microscopy of Membrane Proteins (no. IC200100052). This work was funded in part by an NHMRC Program Grant (no. 1150083) to P.M.S. High-performance computing was supported by Monash MASSIVE. Cryo-EM samples were imaged at the Bio21 Advanced Microscopy Facility (The University of Melbourne) and at the Monash Ramaciotti Centre for Cryo-Electron Microscopy. The authors thank Dr. Hari Venugopal for assistance with 300 kV data collection at the Monash Ramaciotti Centre. B.P.C. P.M.S. and D.W. contributed to conceptualization. B.P.C. expressed protein and purified complexes. Y.J. and I.C.R. prepared baculovirus. E.J.G. and M.M.F. performed assays. B.P.C. and M.J.B. prepared cryo-EM samples and acquired EM data. B.P.C. M.J.B. and S.J.P. processed single-particle cryo-EM data. B.P.C. built atomic models. B.P.C. E.J.G. and S.J.P. prepared figures. All authors reviewed and/or revised the paper. P.M.S. is a co-founder and shareholder of Septerna Inc. D.W. is a shareholder of Septerna Inc. Publisher Copyright: {\textcopyright} 2023 Elsevier Ltd",

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doi = "10.1016/j.str.2023.04.002",

language = "English",

volume = "31",

pages = "668--676.e5",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "6",

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T1 - Molecular insights into peptide agonist engagement with the PTH receptor

AU - Cary, Brian P.

AU - Gerrard, Elliot J.

AU - Belousoff, Matthew J.

AU - Fletcher, Madeleine M.

AU - Jiang, Yan

AU - Russell, Isabella C.

AU - Piper, Sarah J.

AU - Wootten, Denise

AU - Sexton, Patrick M.

N1 - Funding Information: P.M.S. is a Senior Principal Research Fellow (no. 1154434) and D.W. a Senior Research Fellow (no. 1155302) of the National Health and Medical Research Council of Australia. P.M.S. is the Director and D.W. the Monash University Node leader of the Australian Research Council Industrial Transformation Training Centre for Cryo-electron Microscopy of Membrane Proteins (no. IC200100052). This work was funded in part by an NHMRC Program Grant (no. 1150083 ) to P.M.S. High-performance computing was supported by Monash MASSIVE. Cryo-EM samples were imaged at the Bio21 Advanced Microscopy Facility (The University of Melbourne) and at the Monash Ramaciotti Centre for Cryo-Electron Microscopy. The authors thank Dr. Hari Venugopal for assistance with 300 kV data collection at the Monash Ramaciotti Centre. Funding Information: P.M.S. is a Senior Principal Research Fellow (no. 1154434) and D.W. a Senior Research Fellow (no. 1155302) of the National Health and Medical Research Council of Australia. P.M.S. is the Director and D.W. the Monash University Node leader of the Australian Research Council Industrial Transformation Training Centre for Cryo-electron Microscopy of Membrane Proteins (no. IC200100052). This work was funded in part by an NHMRC Program Grant (no. 1150083) to P.M.S. High-performance computing was supported by Monash MASSIVE. Cryo-EM samples were imaged at the Bio21 Advanced Microscopy Facility (The University of Melbourne) and at the Monash Ramaciotti Centre for Cryo-Electron Microscopy. The authors thank Dr. Hari Venugopal for assistance with 300 kV data collection at the Monash Ramaciotti Centre. B.P.C. P.M.S. and D.W. contributed to conceptualization. B.P.C. expressed protein and purified complexes. Y.J. and I.C.R. prepared baculovirus. E.J.G. and M.M.F. performed assays. B.P.C. and M.J.B. prepared cryo-EM samples and acquired EM data. B.P.C. M.J.B. and S.J.P. processed single-particle cryo-EM data. B.P.C. built atomic models. B.P.C. E.J.G. and S.J.P. prepared figures. All authors reviewed and/or revised the paper. P.M.S. is a co-founder and shareholder of Septerna Inc. D.W. is a shareholder of Septerna Inc. Publisher Copyright: © 2023 Elsevier Ltd

PY - 2023/6/1

Y1 - 2023/6/1

N2 - The parathyroid hormone (PTH) 1 receptor (PTH1R) is a G protein-coupled receptor (GPCR) that regulates skeletal development and calcium homeostasis. Here, we describe cryo-EM structures of the PTH1R in complex with fragments of the two hormones, PTH and PTH-related protein, the drug abaloparatide, as well as the engineered tool compounds, long-acting PTH (LA-PTH) and the truncated peptide, M-PTH(1–14). We found that the critical N terminus of each agonist engages the transmembrane bundle in a topologically similar fashion, reflecting similarities in measures of Gαs activation. The full-length peptides induce subtly different extracellular domain (ECD) orientations relative to the transmembrane domain. In the structure bound to M-PTH, the ECD is unresolved, demonstrating that the ECD is highly dynamic when unconstrained by a peptide. High resolutions enabled identification of water molecules near peptide and G protein binding sites. Our results illuminate the action of orthosteric agonists of the PTH1R.

AB - The parathyroid hormone (PTH) 1 receptor (PTH1R) is a G protein-coupled receptor (GPCR) that regulates skeletal development and calcium homeostasis. Here, we describe cryo-EM structures of the PTH1R in complex with fragments of the two hormones, PTH and PTH-related protein, the drug abaloparatide, as well as the engineered tool compounds, long-acting PTH (LA-PTH) and the truncated peptide, M-PTH(1–14). We found that the critical N terminus of each agonist engages the transmembrane bundle in a topologically similar fashion, reflecting similarities in measures of Gαs activation. The full-length peptides induce subtly different extracellular domain (ECD) orientations relative to the transmembrane domain. In the structure bound to M-PTH, the ECD is unresolved, demonstrating that the ECD is highly dynamic when unconstrained by a peptide. High resolutions enabled identification of water molecules near peptide and G protein binding sites. Our results illuminate the action of orthosteric agonists of the PTH1R.

KW - Cryo-EM

KW - g protein-coupled receptor

KW - GPCR

KW - hormone

KW - membrane protein

KW - parathyroid

KW - peptides

KW - PTH

UR - https://www.scopus.com/pages/publications/85160233627

U2 - 10.1016/j.str.2023.04.002

DO - 10.1016/j.str.2023.04.002

M3 - Article

C2 - 37148874

AN - SCOPUS:85160233627

SN - 0969-2126

VL - 31

SP - 668-676.e5

JO - Structure

JF - Structure

IS - 6

ER -

Molecular insights into peptide agonist engagement with the PTH receptor

Abstract

Keywords

Access to Document

Other files and links

ARC Industrial Transformation Training Centre for Cryo-Electron Microscopy of Membrane Proteins for Drug Discovery

Novel targeting of the glucagon family of G protein coupled receptors

Translating membrane proteins into therapeutics; from bedside to bench

High-performance Computing (M3/MASSIVE)

Ramaciotti Centre for Cryo-Electron Microscopy (CryoEM)

Cite this

Molecular insights into peptide agonist engagement with the PTH receptor

Abstract

Keywords

Access to Document

Other files and links

Projects

ARC Industrial Transformation Training Centre for Cryo-Electron Microscopy of Membrane Proteins for Drug Discovery

Novel targeting of the glucagon family of G protein coupled receptors

Translating membrane proteins into therapeutics; from bedside to bench

Equipment

High-performance Computing (M3/MASSIVE)

Ramaciotti Centre for Cryo-Electron Microscopy (CryoEM)

Cite this