Apelin-36-[L28A] and Apelin-36-[L28C(30kDa-PEG)] peptides that improve diet induced obesity are G protein biased ligands at the apelin receptor.
Kuc, Rhoda E
Williams, Thomas L
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Nyimanu, D., Kuc, R. E., Williams, T. L., Bednarek, M., Ambery, P., Jermutus, L., Maguire, J., & et al. (2019). Apelin-36-[L28A] and Apelin-36-[L28C(30kDa-PEG)] peptides that improve diet induced obesity are G protein biased ligands at the apelin receptor.. Peptides, 121 170139. https://doi.org/10.1016/j.peptides.2019.170139
Background: Apelin signalling pathways have important cardiovascular and metabolic functions. Recently, apelin-36-[L28A] and apelin-36-[L28C(30kDa-PEG)], were reported to function independent of the apelin receptor in vivo to produce beneficial metabolic effects without modulating blood pressure. We aimed to show that these peptides bound to the apelin receptor and to further characterise their pharmacology in vitro at the human apelin receptor. Methods: [Pyr1]apelin-13 saturation binding experiments and competition binding experiments were performed in rat and human heart homogenates using [125I]apelin-13 (0.1nM), and/or increasing concentrations of apelin-36, apelin-36-[L28A] and apelin-36-[L28C(30kDa-PEG)] (50pM-100µM). Apelin-36 and its analogues apelin-36-[F36A], apelin-36-[L28A], apelin-36-[L28C(30kDa-PEG)], apelin-36-[A28 A13] and [40kDa-PEG]-apelin-36 were tested in forskolin-induced cAMP inhibition and β–arrestin assays in CHO-K1 cells heterologously expressing the human apelin receptor. Bias signaling was quantified using the operational model for bias. Results: In both species, [Pyr1]apelin-13 had comparable subnanomolar affinity and the apelin receptor density was similar. Apelin-36, apelin-36-[L28A] and apelin-36-[L28C(30kDa-PEG)] competed for binding of [125I]apelin-13 with nanomolar affinities. Apelin-36-[L28A] and apelin-36-[L28C(30kDa-PEG)] inhibited forskolin-induced cAMP release, with nanomolar potencies but they were less potent compared to apelin-36 at recruiting β-arrestin. Bias analysis suggested that these peptides were G protein biased. Additionally, [40kDa-PEG]-apelin-36 and apelin-36-[F36A] retained nanomolar potencies in both cAMP and β-arrestin assays whilst apelin-36-[A13 A28] exhibited a similar profile to apelin-36-[L28C(30kDa-PEG)] in the β–arrestin assay but was more potent in the cAMP assay. Conclusions: Apelin-36-[L28A] and apelin-36-[L28C(30kDa-PEG)] are G protein biased ligand apelin receptor, suggesting that the apelin receptor is an important therapeutic target in metabolic diseases.
Heart Ventricles, CHO Cells, Animals, Humans, Cricetulus, Rats, Rats, Sprague-Dawley, Peptides, Cyclic AMP, Complex Mixtures, Ligands, Binding, Competitive, Protein Binding, Adult, Middle Aged, Female, Male, Colforsin, beta-Arrestins, Apelin, Apelin Receptors
Wellcome Trust [WT107715/Z/15/Z ], Wellcome Trust Programme in Metabolic and Cardiovascular Disease [203814/Z/16/A]; Astra-Zeneca, Cambridge Biomedical Research Centre Biomedical Resources Grant, University of Cambridge, Cardiovascular Theme, RG64226.
Cambridge University Hospitals NHS Foundation Trust (CUH) (unknown)
WELLCOME TRUST (107715/Z/15/Z)
Wellcome Trust (203814/Z/16/Z)
External DOI: https://doi.org/10.1016/j.peptides.2019.170139
This record's URL: https://www.repository.cam.ac.uk/handle/1810/296250
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/