Structural and functional determination of peptide versus small molecule ligand binding at the apelin receptor
We describe a structural and functional study of the G protein-coupled apelin receptor, which binds two endogenous peptide ligands, apelin and Elabela/Toddler (ELA), to regulate cardiovascular development and function. Characterisation of naturally occurring apelin receptor variants from the UK Genomics England 100,000 Genomes Project, and AlphaFold2 modelling, identifies T89<sup>2.64</sup> as important in the ELA binding site, and R168<sup>4.64</sup> as forming extensive interactions with the C-termini of both peptides. Base editing to introduce an R/H168<sup>4.64</sup> variant into human stem cell-derived cardiomyocytes demonstrates that this residue is critical for receptor binding and function. Additionally, we present an apelin receptor crystal structure bound to the G protein-biased, small molecule agonist, CMF-019, which reveals a deeper binding mode versus the endogenous peptides at lipophilic pockets between transmembrane helices associated with GPCR activation. Overall, the data provide proof-of-principle for using genetic variation to identify key sites regulating receptor-ligand engagement.
Authors
- Stefan Gräf (ORCID: https://orcid.org/0000-0002-1315-8873)
- Rhoda E. Kuc (ORCID: https://orcid.org/0000-0003-4673-1602)
- Nicolae Solcan
- Janet J. Maguire (ORCID: https://orcid.org/0000-0002-9254-7040)
- Chris de Graaf (ORCID: https://orcid.org/0000-0002-1226-2150)
- Grégory Verdon (ORCID: https://orcid.org/0000-0002-9788-4361)
- Robyn Macrae (ORCID: https://orcid.org/0000-0002-8129-7242)
- Sanjay Sinha (ORCID: https://orcid.org/0000-0001-5900-1209)
- Oliver Schlenker
- A. N. Zhukov (ORCID: https://orcid.org/0000-0002-2542-9810)
- Thomas Williams (ORCID: https://orcid.org/0000-0002-1051-0595)
- Anthony P. Davenport (ORCID: https://orcid.org/0000-0002-2096-3117)
- Heather Currinn (ORCID: https://orcid.org/0000-0003-1314-1868)
- Brian J. Bender (ORCID: https://orcid.org/0000-0001-9251-9480)
- Jason Brown (ORCID: https://orcid.org/0000-0001-7476-9144)
- Marco Schütz
- Alastair Brown
Institutions
- NHS Blood and Transplant (GB)
- Wellcome/MRC Cambridge Stem Cell Institute (GB)
- University of Cambridge (GB)
- Kyocera (United States) (US)
- National Institute for Health and Care Research (GB)
- Heptares Therapeutics (United Kingdom) (GB)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-12-27
- DOI
- https://doi.org/10.1038/s41467-024-55381-w
- Citations
- 8
- Primary Topic
- Apelin-related biomedical research
- Type
- article
- Field-Weighted Citation Impact
- 3.58
Funders
- Wellcome
- NHS Blood and Transplant
- Wellcome Trust
- National Institute for Health and Care Research
- British Heart Foundation
- Department of Health and Social Care
- NIHR BioResource
- Medical Research Council
- NIHR Cambridge Biomedical Research Centre