Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line

Abstract Sweet taste perception is mediated by a heterodimeric receptor formed by the assembly of the TAS1R2 and TAS1R3 subunits. TAS1R2 and TAS1R3 are class C G-protein-coupled receptors whose members share a common topology, including a large extracellular N-terminal domain (NTD) linked to a seven...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Christine Belloir, Marine Brulé, Lucie Tornier, Fabrice Neiers, Loïc Briand
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/55e2fa0649d34f90887a892f3ce58635
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:55e2fa0649d34f90887a892f3ce58635
record_format	dspace
spelling	oai:doaj.org-article:55e2fa0649d34f90887a892f3ce586352021-11-21T12:22:56ZBiophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line10.1038/s41598-021-01731-32045-2322https://doaj.org/article/55e2fa0649d34f90887a892f3ce586352021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01731-3https://doaj.org/toc/2045-2322Abstract Sweet taste perception is mediated by a heterodimeric receptor formed by the assembly of the TAS1R2 and TAS1R3 subunits. TAS1R2 and TAS1R3 are class C G-protein-coupled receptors whose members share a common topology, including a large extracellular N-terminal domain (NTD) linked to a seven transmembrane domain (TMD) by a cysteine-rich domain. TAS1R2-NTD contains the primary binding site for sweet compounds, including natural sugars and high-potency sweeteners, whereas the TAS1R2-TMD has been shown to bind a limited number of sweet tasting compounds. To understand the molecular mechanisms governing receptor–ligand interactions, we overexpressed the human TAS1R2 (hTAS1R2) in a stable tetracycline-inducible HEK293S cell line and purified the detergent-solubilized receptor. Circular dichroism spectroscopic studies revealed that hTAS1R2 was properly folded with evidence of secondary structures. Using size exclusion chromatography coupled to light scattering, we found that the hTAS1R2 subunit is a dimer. Ligand binding properties were quantified by intrinsic tryptophan fluorescence. Due to technical limitations, natural sugars have not been tested. However, we showed that hTAS1R2 is capable of binding high potency sweeteners with K d values that are in agreement with physiological detection. This study offers a new experimental strategy to identify new sweeteners or taste modulators that act on the hTAS1R2 and is a prerequisite for structural query and biophysical studies.Christine BelloirMarine BruléLucie TornierFabrice NeiersLoïc BriandNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christine Belloir Marine Brulé Lucie Tornier Fabrice Neiers Loïc Briand Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
description	Abstract Sweet taste perception is mediated by a heterodimeric receptor formed by the assembly of the TAS1R2 and TAS1R3 subunits. TAS1R2 and TAS1R3 are class C G-protein-coupled receptors whose members share a common topology, including a large extracellular N-terminal domain (NTD) linked to a seven transmembrane domain (TMD) by a cysteine-rich domain. TAS1R2-NTD contains the primary binding site for sweet compounds, including natural sugars and high-potency sweeteners, whereas the TAS1R2-TMD has been shown to bind a limited number of sweet tasting compounds. To understand the molecular mechanisms governing receptor–ligand interactions, we overexpressed the human TAS1R2 (hTAS1R2) in a stable tetracycline-inducible HEK293S cell line and purified the detergent-solubilized receptor. Circular dichroism spectroscopic studies revealed that hTAS1R2 was properly folded with evidence of secondary structures. Using size exclusion chromatography coupled to light scattering, we found that the hTAS1R2 subunit is a dimer. Ligand binding properties were quantified by intrinsic tryptophan fluorescence. Due to technical limitations, natural sugars have not been tested. However, we showed that hTAS1R2 is capable of binding high potency sweeteners with K d values that are in agreement with physiological detection. This study offers a new experimental strategy to identify new sweeteners or taste modulators that act on the hTAS1R2 and is a prerequisite for structural query and biophysical studies.
format	article
author	Christine Belloir Marine Brulé Lucie Tornier Fabrice Neiers Loïc Briand
author_facet	Christine Belloir Marine Brulé Lucie Tornier Fabrice Neiers Loïc Briand
author_sort	Christine Belloir
title	Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
title_short	Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
title_full	Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
title_fullStr	Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
title_full_unstemmed	Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line
title_sort	biophysical and functional characterization of the human tas1r2 sweet taste receptor overexpressed in a hek293s inducible cell line
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/55e2fa0649d34f90887a892f3ce58635
work_keys_str_mv	AT christinebelloir biophysicalandfunctionalcharacterizationofthehumantas1r2sweettastereceptoroverexpressedinahek293sinduciblecellline AT marinebrule biophysicalandfunctionalcharacterizationofthehumantas1r2sweettastereceptoroverexpressedinahek293sinduciblecellline AT lucietornier biophysicalandfunctionalcharacterizationofthehumantas1r2sweettastereceptoroverexpressedinahek293sinduciblecellline AT fabriceneiers biophysicalandfunctionalcharacterizationofthehumantas1r2sweettastereceptoroverexpressedinahek293sinduciblecellline AT loicbriand biophysicalandfunctionalcharacterizationofthehumantas1r2sweettastereceptoroverexpressedinahek293sinduciblecellline
_version_	1718419065216696320

Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line

Ejemplares similares