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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Federation of American Societies for Experimental Biology
2012
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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. |
format | Online Article Text |
id | pubmed-3509056 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Federation of American Societies for Experimental Biology |
record_format | MEDLINE/PubMed |
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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