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An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor
Sweet taste is mediated by a dimeric receptor composed of two distinct subunits, T1R2 and T1R3, whereas the T1R1/T1R3 receptor is involved in umami taste perception. The T1R1, T1R2, and T1R3 subunits are members of the small family of class C G protein-coupled receptors (GPCRs). The members of this...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Landes Bioscience
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3566016/ https://www.ncbi.nlm.nih.gov/pubmed/22909933 http://dx.doi.org/10.4161/bioe.21877 |
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author | Maîtrepierre, Elodie Sigoillot, Maud Le Pessot, Laurence Briand, Loïc |
author_facet | Maîtrepierre, Elodie Sigoillot, Maud Le Pessot, Laurence Briand, Loïc |
author_sort | Maîtrepierre, Elodie |
collection | PubMed |
description | Sweet taste is mediated by a dimeric receptor composed of two distinct subunits, T1R2 and T1R3, whereas the T1R1/T1R3 receptor is involved in umami taste perception. The T1R1, T1R2, and T1R3 subunits are members of the small family of class C G protein-coupled receptors (GPCRs). The members of this family are characterized by a large N-terminal domain (NTD), which is structurally similar to bacterial periplasmic-binding proteins and contains the primary ligand-binding site. In a recent study, we described a strategy to produce a functional dimeric human T1R3-NTD. Although the protein was expressed as inclusion bodies (IBs) using the Escherichia coli system, the conditions for the refolding of functional hT1R3-NTD were determined using a fractional factorial screen coupled to a binding assay. Here, we report that this refolding strategy can be used to produce T1R1- and T1R2-NTDs in large quantities. We also discuss that our findings could be more generally applicable to other class C GPCR-NTDs, including the γ-aminobutyric acid type B receptor (GABA(B)R), the extracellular calcium-sensing receptor (CaSR) and the large family of pheromone (V2R) orphan receptors. |
format | Online Article Text |
id | pubmed-3566016 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Landes Bioscience |
record_format | MEDLINE/PubMed |
spelling | pubmed-35660162013-02-13 An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor Maîtrepierre, Elodie Sigoillot, Maud Le Pessot, Laurence Briand, Loïc Bioengineered Research Note Sweet taste is mediated by a dimeric receptor composed of two distinct subunits, T1R2 and T1R3, whereas the T1R1/T1R3 receptor is involved in umami taste perception. The T1R1, T1R2, and T1R3 subunits are members of the small family of class C G protein-coupled receptors (GPCRs). The members of this family are characterized by a large N-terminal domain (NTD), which is structurally similar to bacterial periplasmic-binding proteins and contains the primary ligand-binding site. In a recent study, we described a strategy to produce a functional dimeric human T1R3-NTD. Although the protein was expressed as inclusion bodies (IBs) using the Escherichia coli system, the conditions for the refolding of functional hT1R3-NTD were determined using a fractional factorial screen coupled to a binding assay. Here, we report that this refolding strategy can be used to produce T1R1- and T1R2-NTDs in large quantities. We also discuss that our findings could be more generally applicable to other class C GPCR-NTDs, including the γ-aminobutyric acid type B receptor (GABA(B)R), the extracellular calcium-sensing receptor (CaSR) and the large family of pheromone (V2R) orphan receptors. Landes Bioscience 2013-01-01 /pmc/articles/PMC3566016/ /pubmed/22909933 http://dx.doi.org/10.4161/bioe.21877 Text en Copyright © 2012 Landes Bioscience http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited. |
spellingShingle | Research Note Maîtrepierre, Elodie Sigoillot, Maud Le Pessot, Laurence Briand, Loïc An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title_full | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title_fullStr | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title_full_unstemmed | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title_short | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
title_sort | efficient escherichia coli expression system for the production of a functional n-terminal domain of the t1r3 taste receptor |
topic | Research Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3566016/ https://www.ncbi.nlm.nih.gov/pubmed/22909933 http://dx.doi.org/10.4161/bioe.21877 |
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