A novel functional screening assay to monitor sweet taste receptor activation in vitro

Shanna Bastiaan-Net, Dianne B.P.M. van den Berg-Somhorst, Renata M.C. Ariëns, Marcel Paques, Jurriaan J. Mes

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The human sweet taste receptor is a heterodimer comprised of the class C G protein-coupled receptor (GPCR) subunits TAS1R2 and TAS1R3. A wide collection of sweet tasting compounds and modulators of sweet taste interact with this receptor. Although TAS1R2/TAS1R3-mediated signaling is well-studied, the molecular basis for its desensitization remains unclear while such knowledge would signify a profound step forward in understanding the mechanism behind sweet taste perception and taste modulation. In this work, the possible involvement of β-arrestin in downstream signaling was investigated. A stable clonal Human Embryonic Kidney (HEK)-derived cell line containing the PathHunter™ GPCR technology was developed, in which β-arrestin-mediated endosomal receptor internalization can be monitored by ligand-induced enzyme complementation of β-galactosidase (β-gal). Stimulatory responses and antibody-specific receptor detection indicated that the TAS1R2/TAS1R3 receptor is endogenously expressed in this clonal cell line. Natural sugars (including fructose, glucose, sucrose, maltose, maltitol and mannitol) and artificial sweeteners (acesulfame-K and sucralose) stimulated enzyme complementation activity in a concentration dependent manner. Besides, we observed that the assay detected modification of sugar induced cell responses by sweetness enhancers. These results combined implicate that TAS1R2/TAS1R3 receptor desensitization by internalization is most likely mediated by β-arrestin-induced endocytosis. This assay approach, making use of naturally expressed TAS1R2/TAS1R3 receptors and required co-factors, further allows effective screening for and development of novel high potency non-caloric sweeteners, sweet taste modulators or optimal blends with enhanced sweet taste.
LanguageEnglish
Pages173-183
JournalFlavour and Fragrance Journal
Volume33
Issue number2
Early online date21 Nov 2017
DOIs
Publication statusPublished - 18 Feb 2018

Fingerprint

Arrestin
Sweetening Agents
Assays
Screening
trichlorosucrose
Chemical activation
arrestins
G-Protein-Coupled Receptors
screening
Sugars
Modulators
receptors
monitoring
assays
Cells
Maltose
Enzyme activity
Mannitol
beta-Galactosidase
Fructose

Keywords

  • G protein-coupled receptor (GPCR)
  • Sweet modulators
  • Sweet taste receptor
  • TAS1R2-TAS1R3
  • β-arrestin

Cite this

@article{fafd157d819f4e118d365e5ac32ef254,
title = "A novel functional screening assay to monitor sweet taste receptor activation in vitro",
abstract = "The human sweet taste receptor is a heterodimer comprised of the class C G protein-coupled receptor (GPCR) subunits TAS1R2 and TAS1R3. A wide collection of sweet tasting compounds and modulators of sweet taste interact with this receptor. Although TAS1R2/TAS1R3-mediated signaling is well-studied, the molecular basis for its desensitization remains unclear while such knowledge would signify a profound step forward in understanding the mechanism behind sweet taste perception and taste modulation. In this work, the possible involvement of β-arrestin in downstream signaling was investigated. A stable clonal Human Embryonic Kidney (HEK)-derived cell line containing the PathHunter™ GPCR technology was developed, in which β-arrestin-mediated endosomal receptor internalization can be monitored by ligand-induced enzyme complementation of β-galactosidase (β-gal). Stimulatory responses and antibody-specific receptor detection indicated that the TAS1R2/TAS1R3 receptor is endogenously expressed in this clonal cell line. Natural sugars (including fructose, glucose, sucrose, maltose, maltitol and mannitol) and artificial sweeteners (acesulfame-K and sucralose) stimulated enzyme complementation activity in a concentration dependent manner. Besides, we observed that the assay detected modification of sugar induced cell responses by sweetness enhancers. These results combined implicate that TAS1R2/TAS1R3 receptor desensitization by internalization is most likely mediated by β-arrestin-induced endocytosis. This assay approach, making use of naturally expressed TAS1R2/TAS1R3 receptors and required co-factors, further allows effective screening for and development of novel high potency non-caloric sweeteners, sweet taste modulators or optimal blends with enhanced sweet taste.",
keywords = "G protein-coupled receptor (GPCR), Sweet modulators, Sweet taste receptor, TAS1R2-TAS1R3, β-arrestin",
author = "Shanna Bastiaan-Net and {van den Berg-Somhorst}, {Dianne B.P.M.} and Ari{\"e}ns, {Renata M.C.} and Marcel Paques and Mes, {Jurriaan J.}",
year = "2018",
month = "2",
day = "18",
doi = "10.1002/ffj.3431",
language = "English",
volume = "33",
pages = "173--183",
journal = "Flavour and Fragrance Journal",
issn = "0882-5734",
publisher = "Wiley",
number = "2",

}

A novel functional screening assay to monitor sweet taste receptor activation in vitro. / Bastiaan-Net, Shanna; van den Berg-Somhorst, Dianne B.P.M.; Ariëns, Renata M.C.; Paques, Marcel; Mes, Jurriaan J.

In: Flavour and Fragrance Journal, Vol. 33, No. 2, 18.02.2018, p. 173-183.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - A novel functional screening assay to monitor sweet taste receptor activation in vitro

AU - Bastiaan-Net, Shanna

AU - van den Berg-Somhorst, Dianne B.P.M.

AU - Ariëns, Renata M.C.

AU - Paques, Marcel

AU - Mes, Jurriaan J.

PY - 2018/2/18

Y1 - 2018/2/18

N2 - The human sweet taste receptor is a heterodimer comprised of the class C G protein-coupled receptor (GPCR) subunits TAS1R2 and TAS1R3. A wide collection of sweet tasting compounds and modulators of sweet taste interact with this receptor. Although TAS1R2/TAS1R3-mediated signaling is well-studied, the molecular basis for its desensitization remains unclear while such knowledge would signify a profound step forward in understanding the mechanism behind sweet taste perception and taste modulation. In this work, the possible involvement of β-arrestin in downstream signaling was investigated. A stable clonal Human Embryonic Kidney (HEK)-derived cell line containing the PathHunter™ GPCR technology was developed, in which β-arrestin-mediated endosomal receptor internalization can be monitored by ligand-induced enzyme complementation of β-galactosidase (β-gal). Stimulatory responses and antibody-specific receptor detection indicated that the TAS1R2/TAS1R3 receptor is endogenously expressed in this clonal cell line. Natural sugars (including fructose, glucose, sucrose, maltose, maltitol and mannitol) and artificial sweeteners (acesulfame-K and sucralose) stimulated enzyme complementation activity in a concentration dependent manner. Besides, we observed that the assay detected modification of sugar induced cell responses by sweetness enhancers. These results combined implicate that TAS1R2/TAS1R3 receptor desensitization by internalization is most likely mediated by β-arrestin-induced endocytosis. This assay approach, making use of naturally expressed TAS1R2/TAS1R3 receptors and required co-factors, further allows effective screening for and development of novel high potency non-caloric sweeteners, sweet taste modulators or optimal blends with enhanced sweet taste.

AB - The human sweet taste receptor is a heterodimer comprised of the class C G protein-coupled receptor (GPCR) subunits TAS1R2 and TAS1R3. A wide collection of sweet tasting compounds and modulators of sweet taste interact with this receptor. Although TAS1R2/TAS1R3-mediated signaling is well-studied, the molecular basis for its desensitization remains unclear while such knowledge would signify a profound step forward in understanding the mechanism behind sweet taste perception and taste modulation. In this work, the possible involvement of β-arrestin in downstream signaling was investigated. A stable clonal Human Embryonic Kidney (HEK)-derived cell line containing the PathHunter™ GPCR technology was developed, in which β-arrestin-mediated endosomal receptor internalization can be monitored by ligand-induced enzyme complementation of β-galactosidase (β-gal). Stimulatory responses and antibody-specific receptor detection indicated that the TAS1R2/TAS1R3 receptor is endogenously expressed in this clonal cell line. Natural sugars (including fructose, glucose, sucrose, maltose, maltitol and mannitol) and artificial sweeteners (acesulfame-K and sucralose) stimulated enzyme complementation activity in a concentration dependent manner. Besides, we observed that the assay detected modification of sugar induced cell responses by sweetness enhancers. These results combined implicate that TAS1R2/TAS1R3 receptor desensitization by internalization is most likely mediated by β-arrestin-induced endocytosis. This assay approach, making use of naturally expressed TAS1R2/TAS1R3 receptors and required co-factors, further allows effective screening for and development of novel high potency non-caloric sweeteners, sweet taste modulators or optimal blends with enhanced sweet taste.

KW - G protein-coupled receptor (GPCR)

KW - Sweet modulators

KW - Sweet taste receptor

KW - TAS1R2-TAS1R3

KW - β-arrestin

U2 - 10.1002/ffj.3431

DO - 10.1002/ffj.3431

M3 - Article

VL - 33

SP - 173

EP - 183

JO - Flavour and Fragrance Journal

T2 - Flavour and Fragrance Journal

JF - Flavour and Fragrance Journal

SN - 0882-5734

IS - 2

ER -