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Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs

When it is difficult to develop selective ligands within a family of related G-protein-coupled receptors (GPCRs), chemically engineered receptors activated solely by synthetic ligands (RASSLs) are useful alternatives for probing receptor function. In the present work, we explored whether a RASSL of...

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Autores principales: Hudson, Brian D., Christiansen, Elisabeth, Tikhonova, Irina G., Grundmann, Manuel, Kostenis, Evi, Adams, David R., Ulven, Trond, Milligan, Graeme
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Federation of American Societies for Experimental Biology 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3509056/
https://www.ncbi.nlm.nih.gov/pubmed/22919070
http://dx.doi.org/10.1096/fj.12-213314
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author Hudson, Brian D.
Christiansen, Elisabeth
Tikhonova, Irina G.
Grundmann, Manuel
Kostenis, Evi
Adams, David R.
Ulven, Trond
Milligan, Graeme
author_facet Hudson, Brian D.
Christiansen, Elisabeth
Tikhonova, Irina G.
Grundmann, Manuel
Kostenis, Evi
Adams, David R.
Ulven, Trond
Milligan, Graeme
author_sort Hudson, Brian D.
collection PubMed
description When it is difficult to develop selective ligands within a family of related G-protein-coupled receptors (GPCRs), chemically engineered receptors activated solely by synthetic ligands (RASSLs) are useful alternatives for probing receptor function. In the present work, we explored whether a RASSL of the free fatty acid receptor 2 (FFA2) could be developed on the basis of pharmacological variation between species orthologs. For this, bovine FFA2 was characterized, revealing distinct ligand selectivity compared with human FFA2. Homology modeling and mutational analysis demonstrated a single mutation in human FFA2 of C4.57G resulted in a human FFA2 receptor with ligand selectivity similar to the bovine receptor. This was exploited to generate human FFA2-RASSL by the addition of a second mutation at a known orthosteric ligand interaction site, H6.55Q. The resulting FFA2-RASSL displayed a >100-fold loss of activity to endogenous ligands, while responding to the distinct ligand sorbic acid with pEC(50) values for inhibition of cAMP, 5.83 ± 0.11; Ca(2+) mobilization, 4.63 ± 0.05; ERK phosphorylation, 5.61 ± 0.06; and dynamic mass redistribution, 5.35 ± 0.06. This FFA2-RASSL will be useful in future studies on this receptor and demonstrates that exploitation of pharmacological variation between species orthologs is a powerful method to generate novel chemically engineered GPCRs.—Hudson, B. D., Christiansen, E., Tikhonova, I. G., Grundmann, M., Kostenis, E., Adams, D. R., Ulven, T., Milligan, G. Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs.
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spelling pubmed-35090562012-12-13 Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs Hudson, Brian D. Christiansen, Elisabeth Tikhonova, Irina G. Grundmann, Manuel Kostenis, Evi Adams, David R. Ulven, Trond Milligan, Graeme FASEB J Research Communications When it is difficult to develop selective ligands within a family of related G-protein-coupled receptors (GPCRs), chemically engineered receptors activated solely by synthetic ligands (RASSLs) are useful alternatives for probing receptor function. In the present work, we explored whether a RASSL of the free fatty acid receptor 2 (FFA2) could be developed on the basis of pharmacological variation between species orthologs. For this, bovine FFA2 was characterized, revealing distinct ligand selectivity compared with human FFA2. Homology modeling and mutational analysis demonstrated a single mutation in human FFA2 of C4.57G resulted in a human FFA2 receptor with ligand selectivity similar to the bovine receptor. This was exploited to generate human FFA2-RASSL by the addition of a second mutation at a known orthosteric ligand interaction site, H6.55Q. The resulting FFA2-RASSL displayed a >100-fold loss of activity to endogenous ligands, while responding to the distinct ligand sorbic acid with pEC(50) values for inhibition of cAMP, 5.83 ± 0.11; Ca(2+) mobilization, 4.63 ± 0.05; ERK phosphorylation, 5.61 ± 0.06; and dynamic mass redistribution, 5.35 ± 0.06. This FFA2-RASSL will be useful in future studies on this receptor and demonstrates that exploitation of pharmacological variation between species orthologs is a powerful method to generate novel chemically engineered GPCRs.—Hudson, B. D., Christiansen, E., Tikhonova, I. G., Grundmann, M., Kostenis, E., Adams, D. R., Ulven, T., Milligan, G. Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs. Federation of American Societies for Experimental Biology 2012-12 /pmc/articles/PMC3509056/ /pubmed/22919070 http://dx.doi.org/10.1096/fj.12-213314 Text en © FASEB This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/us/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Communications
Hudson, Brian D.
Christiansen, Elisabeth
Tikhonova, Irina G.
Grundmann, Manuel
Kostenis, Evi
Adams, David R.
Ulven, Trond
Milligan, Graeme
Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title_full Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title_fullStr Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title_full_unstemmed Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title_short Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
title_sort chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs
topic Research Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3509056/
https://www.ncbi.nlm.nih.gov/pubmed/22919070
http://dx.doi.org/10.1096/fj.12-213314
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