Structure–activity relationship of GPR15L peptide analogues and investigation of their interaction with the GPR15 receptor
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The 57-mer full-length GPR15L(25-81) peptide has been identified as the principal endogenous agonist of the G protein-coupled receptor GPR15. Its main activity resides in the C-terminal 11-mer GPR15L(71-81), which has full efficacy but ~40-fold lower potency than the full-length peptide. Here, we systematically investigated the structure–activity relationship of GPR15L(71-81) by truncations/extensions, alanine-scanning, and N- and C-terminal capping. The synthesized peptide analogues were tested at GPR15 stably expressed in HEK293A cells using a homogenous time-resolved Förster resonance energy transfer-based Gi cAMP functional assay. We show that the C-terminal α carboxyl group and the residues Leu78, Pro75, Val74, and Trp72 are critical for receptor interaction and contribute significantly to the peptide potency. Furthermore, we tested the ability of GPR15L(71-81), C-terminally amidated GPR15L(71-81), and GPR15L(25-81) to activate the three GPR15 receptor mutants in a bioluminescence resonance energy transfer-based G protein activation assay. The results demonstrate that the Lys192 and Glu272 residues in GPR15 are important for the potency of the GPR15L peptide. Overall, our study identifies critical residues in the peptide and receptor sequences for future drug design.
Originalsprog | Engelsk |
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Tidsskrift | Basic and Clinical Pharmacology and Toxicology |
Vol/bind | 132 |
Udgave nummer | 6 |
Sider (fra-til) | 459-471 |
ISSN | 1742-7835 |
DOI | |
Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
We thank Asmita Manandhar for performing chemical analyses on some of the synthesized peptides. This project is funded by the China Scholarship Council (Grant 201907940002 to Y.D.), the Novo Nordisk Foundation (Grants NNF17OC0028412 and NNF19OC0057730 to H.B.‐O.), the Carlsberg Foundation (Grant CF20‐0248 to H.B.‐O.), Lundbeck Foundation (Grant R163‐2013‐16327 to D.E.G.), European Research Council (Grant 639125 to D.E.G.) and Novo Nordisk A/S (STAR postdoc program grant to D.E.G.).
Publisher Copyright:
© 2023 The Authors. Basic & Clinical Pharmacology & Toxicology published by John Wiley & Sons Ltd on behalf of Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).
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