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Molecular recognition of two endogenous hormones by the human parathyroid hormone receptor-1

Acta Pharmacologica Sinica volume 44, pages 1227–1237 (2023)Cite this article

Abstract

Parathyroid hormone (PTH) and PTH-related peptide (PTHrP) are two endogenous hormones recognized by PTH receptor-1 (PTH1R), a member of class B G protein- coupled receptors (GPCRs). Both PTH and PTHrP analogs including teriparatide and abaloparatide are approved drugs for osteoporosis, but they exhibit distinct pharmacology. Here we report two cryo-EM structures of human PTH1R bound to PTH and PTHrP in the G protein-bound state at resolutions of 2.62 Å and 3.25 Å, respectively. Detailed analysis of these structures uncovers both common and unique features for the agonism of PTH and PTHrP. Molecular dynamics (MD) simulation together with site-directed mutagenesis studies reveal the molecular basis of endogenous hormones recognition specificity and selectivity to PTH1R. These results provide a rational template for the clinical use of PTH and PTHrP analogs as an anabolic therapy for osteoporosis and other disorders.

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Fig. 1: The overall cryo-EM structures of Gs-coupled PTH1R bound to PTH and PTHrP.
Fig. 2: Structure comparisons of PTH1R-Gs complexes by different ligands.
Fig. 3: Molecular recognition of PTH1–34 and PTHrP1–36 by PTH1R.
Fig. 4: G protein coupling of multi-agonist-bound PTH1R.
Fig. 5: Structures of the active PTH1R in G protein-bound state and comparison with structure of the active PTH1R in the absence of G protein.

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Data availability

Cryo-EM maps have been deposited in the Electron Microscopy Data Bank under accession codes: EMD-34585 (PTH-bound PTH1R-Gs complex), EMD-34587 (PTHrP-bound PTH1R-Gs complex) and EMD-34598 (dimer of PTH-bound PTH1R-Gs complex). The atomic coordinates have been deposited in the Protein Data Bank under accession codes: 8HA0 (PTH-bound PTH1R-Gs complex), 8HAF (PTHrP-bound PTH1R-Gs complex) and 8HAO (dimer of PTH-bound PTH1R-Gs complex).

References

  1. Gardella TJ, Vilardaga JP. International Union of Basic and Clinical Pharmacology. XCIII. The parathyroid hormone receptors–family B G protein-coupled receptors. Pharmacol Rev. 2015;67:310–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Zhao LH, Ma S, Sutkeviciute I, Shen DD, Zhou XE, de Waal PW, et al. Structure and dynamics of the active human parathyroid hormone receptor-1. Science. 2019;364:148–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Nishimura Y, Esaki T, Isshiki Y, Furuta Y, Mizutani A, Kotake T, et al. Lead optimization and avoidance of reactive metabolite leading to PCO371, a potent, selective, and orally available human parathyroid hormone receptor 1 (hPTHR1) agonist. J Med Chem. 2020;63:5089–99.

    Article  CAS  PubMed  Google Scholar 

  4. Kir S, White JP, Kleiner S, Kazak L, Cohen P, Baracos VE, et al. Tumour-derived PTH-related protein triggers adipose tissue browning and cancer cachexia. Nature. 2014;513:100–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Wider J, Undyala VVR, Lanske B, Datta NS, Przyklenk K. Parathyroid hormone-related peptide and its analog, abaloparatide, attenuate lethal myocardial ischemia-reperfusion injury. J Clin Med. 2022;11:2273; https://doi.org/10.3390/jcm11092273.

  6. Chew CK, Clarke BL. Abaloparatide: recombinant human PTHrP (1-34) anabolic therapy for osteoporosis. Maturitas. 2017;97:53–60.

    Article  CAS  PubMed  Google Scholar 

  7. Srivastava M, Deal C. Osteoporosis in elderly: prevention and treatment. Clin Geriatr Med. 2002;18:529–55.

    Article  PubMed  Google Scholar 

  8. Bilezikian JP, Rubin MR, Finkelstein JS. Parathyroid hormone as an anabolic therapy for women and men. J Endocrinol Invest. 2005;28:41–9.

    CAS  PubMed  Google Scholar 

  9. Quattrocchi E, Kourlas H. Teriparatide: a review. Clin Ther. 2004;26:841–54.

    Article  CAS  PubMed  Google Scholar 

  10. Gonnelli S, Caffarelli C. Abaloparatide. Clin Cases Min Bone Metab. 2016;13:106–9.

    Google Scholar 

  11. Pioszak AA, Parker NR, Gardella TJ, Xu HE. Structural basis for parathyroid hormone-related protein binding to the parathyroid hormone receptor and design of conformation-selective peptides. J Biol Chem. 2009;284:28382–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Dean T, Vilardaga JP, Potts JT Jr., Gardella TJ. Altered selectivity of parathyroid hormone (PTH) and PTH-related protein (PTHrP) for distinct conformations of the PTH/PTHrP receptor. Mol Endocrinol. 2008;22:156–66.

    Article  CAS  PubMed  Google Scholar 

  13. Hattersley G, Dean T, Corbin BA, Bahar H, Gardella TJ. Binding selectivity of abaloparatide for PTH-Type-1-receptor conformations and effects on downstream signaling. Endocrinology. 2016;157:141–9.

    Article  CAS  PubMed  Google Scholar 

  14. Okazaki M, Ferrandon S, Vilardaga JP, Bouxsein ML, Potts JT Jr., Gardella TJ. Prolonged signaling at the parathyroid hormone receptor by peptide ligands targeted to a specific receptor conformation. Proc Natl Acad Sci USA. 2008;105:16525–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zhao LH, Lin J, Ji SY, Zhou XE, Mao C, Shen DD, et al. Structure insights into selective coupling of G protein subtypes by a class B G protein-coupled receptor. Nat Commun. 2022;13:6670.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Kobayashi K, Kawakami K, Kusakizako T, Miyauchi H, Tomita A, Kobayashi K, et al. Endogenous ligand recognition and structural transition of a human PTH receptor. Mol Cell. 2022;82:3468–83. e5

    Article  CAS  PubMed  Google Scholar 

  17. Maeda S, Koehl A, Matile H, Hu H, Hilger D, Schertler GFX, et al. Development of an antibody fragment that stabilizes GPCR/G-protein complexes. Nat Commun. 2018;9:3712.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Chan P, Gabay M, Wright FA, Kan W, Oner SS, Lanier SM, et al. Purification of heterotrimeric G protein alpha subunits by GST-Ric-8 association: primary characterization of purified G alpha(olf). J Biol Chem. 2011;286:2625–35.

    Article  CAS  PubMed  Google Scholar 

  19. Dixon AS, Schwinn MK, Hall MP, Zimmerman K, Otto P, Lubben TH, et al. NanoLuc complementation reporter optimized for accurate measurement of protein interactions in cells. ACS Chem Biol. 2016;11:400–8.

    Article  CAS  PubMed  Google Scholar 

  20. Ma S, Shen Q, Zhao LH, Mao C, Zhou XE, Shen DD, et al. Molecular basis for hormone recognition and activation of corticotropin-releasing factor receptors. Mol Cell. 2020;77:669–80 e4.

    Article  CAS  PubMed  Google Scholar 

  21. Zivanov J, Nakane T, Scheres SHW. Estimation of high-order aberrations and anisotropic magnification from cryo-EM data sets in RELION-3.1. Iucrj. 2020;7:253–67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Rohou A, Grigorieff N. CTFFIND4: fast and accurate defocus estimation from electron micrographs. J Struct Biol. 2015;192:216–21.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Punjani A, Rubinstein JL, Fleet DJ, Brubaker MA. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat Methods. 2017;14:290–6.

    Article  CAS  PubMed  Google Scholar 

  24. Sanchez-Garcia R, Gomez-Blanco J, Cuervo A, Carazo JM, Sorzano COS, Vargas J. DeepEMhancer: a deep learning solution for cryo-EM volume post-processing. Commun Biol. 2021;4:874.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, et al. UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem. 2004;25:1605–12.

    Article  CAS  PubMed  Google Scholar 

  26. Emsley P, Cowtan K. Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol Crystallogr. 2004;60:2126–32.

    Article  PubMed  Google Scholar 

  27. Adams PD, Afonine PV, Bunkoczi G, Chen VB, Davis IW, Echols N, et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr. 2010;66:213–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Chen VB, Arendall WB, Headd JJ, Keedy DA, Immormino RM, Kapral GJ, et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr D. 2010;66:12–21.

    Article  CAS  PubMed  Google Scholar 

  29. Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O, et al. Highly accurate protein structure prediction with AlphaFold. Nature. 2021;596:583–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Jo S, Cheng X, Lee J, Kim S, Park S-J, Patel DS, et al. CHARMM-GUI 10 years for biomolecular modeling and simulation. J Comput Chem. 2017;38:1114–24.

    Article  CAS  PubMed  Google Scholar 

  31. Lee J, Hitzenberger M, Rieger M, Kern NR, Zacharias M, Im W. CHARMM-GUI supports the Amber force fields. J Chem Phys. 2020;153:35103.

    Article  CAS  Google Scholar 

  32. Tian C, Kasavajhala K, Belfon KAA, Raguette L, Huang H, Migues AN, et al. ff19SB: amino-acid-specific protein backbone parameters trained against quantum mechanics energy surfaces in solution. J Chem Theory Comput. 2020;16:528–52.

    Article  PubMed  Google Scholar 

  33. He X, Man VH, Yang W, Lee TS, Wang J. A fast and high-quality charge model for the next generation general AMBER force field. J Chem Phys. 2020;153:114502.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Roe DR, Cheatham TE III. PTRAJ and CPPTRAJ: software for processing and analysis of molecular dynamics trajectory data. J Chem theory Comput. 2013;9:3084–95.

    Article  CAS  PubMed  Google Scholar 

  35. Harrigan MP, Sultan MM, Hernández CX, Husic BE, Eastman P, Schwantes CR, et al. MSMBuilder: statistical models for biomolecular dynamics. Biophys J. 2017;112:10–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Miller BR 3rd, McGee TD Jr., Swails JM, Homeyer N, Gohlke H, Roitberg AE. MMPBSA.py: an efficient program for end-state free energy calculations. J Chem Theory Comput. 2012;8:3314–21.

    Article  CAS  PubMed  Google Scholar 

  37. Pioszak AA, Xu HE. Molecular recognition of parathyroid hormone by its G protein-coupled receptor. Proc Natl Acad Sci USA. 2008;105:5034–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Ehrenmann J, Schoppe J, Klenk C, Pluckthun A. New views into class B GPCRs from the crystal structure of PTH1R. FEBS J. 2019;286:4852–60.

    Article  CAS  PubMed  Google Scholar 

  39. Ehrenmann J, Schoppe J, Klenk C, Rappas M, Kummer L, Dore AS, et al. High-resolution crystal structure of parathyroid hormone 1 receptor in complex with a peptide agonist. Nat Struct Mol Biol. 2018;25:1086–92.

    Article  CAS  PubMed  Google Scholar 

  40. Josephs TM, Belousoff MJ, Liang YL, Piper SJ, Cao J, Garama DJ, et al. Structure and dynamics of the CGRP receptor in apo and peptide-bound forms. Science. 2021;372:eabf7258. https://doi.org/10.1126/science.abf7258.

  41. Mannstadt M, Clarke BL, Vokes T, Brandi ML, Ranganath L, Fraser WD, et al. Efficacy and safety of recombinant human parathyroid hormone (1-84) in hypoparathyroidism (REPLACE): a double-blind, placebo-controlled, randomised, phase 3 study. Lancet Diabetes Endocrinol. 2013;1:275–83.

    Article  CAS  PubMed  Google Scholar 

  42. Zhai X, Mao C, Shen Q, Zang S, Shen DD, Zhang H, et al. Molecular insights into the distinct signaling duration for the peptide-induced PTH1R activation. Nat Commun. 2022;13:6276.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Cary BP, Gerrard EJ, Belousoff MJ, Fletcher MM, Jiang Y, Russell IC, et al. Molecular insights into peptide agonist engagement with the PTH1 receptor. Biorxiv. 2022. https://doi.org/10.1101/2022.09.04.506565.

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Acknowledgements

The cryo-EM data were collected by Wen Hu and Kai Wu at Advanced Center for Electron Microscopy at Shanghai Institute of Materia Medica, Chinese Academy of Sciences. We are grateful to them for collecting the cryo-EM data. This work was supported by National Natural Science Foundation of China (32071203 to LHZ; 82073904 to MWW and 81973373 to DHY), the National Key R&D Program of China (2019YFA0904200), the Young Innovator Association of CAS (2018325 to LHZ) and SA-SIBS Scholarship Program to LHZ and DHY; Ministry of Science and Technology (China) grants (2018YFA0507002 to HEX and 2018YFA0507000 to MWW), the Shanghai Municipal Science and Technology Major Project (2019SHZDZX02 to HEX; 18ZR1447800 to LHZ and 21JC1401600 to DHY), the CAS Strategic Priority Research Program (XDB08020303 to HEX).

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Author notes

  1. These authors contributed equally: Li-hua Zhao, Qing-ning Yuan, An-tao Dai

Authors and Affiliations

  1. The CAS Key Laboratory of Receptor Research, Center for Structure and Function of Drug Targets, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Li-hua Zhao, Qing-ning Yuan, An-tao Dai, Xin-heng He, You-wei Xu, Yan Zhou, De-hua Yang & H. Eric Xu

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Li-hua Zhao, Xin-heng He, De-hua Yang & H. Eric Xu

  3. The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    An-tao Dai, Yan Zhou & De-hua Yang

  4. Research Center for Deepsea Bioresources, Sanya, 572025, China

    Chuan-wei Chen & Ming-wei Wang

  5. School of Pharmacy, Fudan University, Shanghai, 201203, China

    Chao Zhang

  6. Department of Pharmacology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China

    Ming-wei Wang

  7. Department of Chemistry, School of Science, The University of Tokyo, Tokyo, 113-0033, Japan

    Ming-wei Wang

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Contributions

LHZ designed the expression constructs, purified the complexes, prepared the final samples for negative stain and data collection toward the structures, participated in model building and performed structure and function data analysis, prepared figures and wrote the manuscript; LHZ prepared the cryo-EM grids and QNY collected cryo-EM images and performed map calculations, built and refined the structure models; YWX participated in model refinement; XHH conducted MD simulations; ATD, CWC, CZ and YZ performed signaling experiments under the supervision of DHY and MWW; LHZ and HEX conceived the project, wrote the manuscript.

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Correspondence to Li-hua Zhao, De-hua Yang or H. Eric Xu.

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Zhao, Lh., Yuan, Qn., Dai, At. et al. Molecular recognition of two endogenous hormones by the human parathyroid hormone receptor-1. Acta Pharmacol Sin 44, 1227–1237 (2023). https://doi.org/10.1038/s41401-022-01032-z

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