Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line.
Cell Line
Chemical Phenomena
Gene Expression
Gene Knock-In Techniques
HEK293 Cells
Humans
Immunohistochemistry
Ligands
Membrane Proteins
/ chemistry
Protein Binding
Protein Conformation
Protein Multimerization
Receptors, G-Protein-Coupled
/ chemistry
Structure-Activity Relationship
Sweetening Agents
/ chemistry
Taste Buds
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
15 11 2021
15 11 2021
Historique:
received:
28
05
2021
accepted:
01
11
2021
entrez:
16
11
2021
pubmed:
17
11
2021
medline:
1
2
2022
Statut:
epublish
Résumé
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
Identifiants
pubmed: 34782704
doi: 10.1038/s41598-021-01731-3
pii: 10.1038/s41598-021-01731-3
pmc: PMC8593021
doi:
Substances chimiques
Ligands
0
Membrane Proteins
0
Receptors, G-Protein-Coupled
0
Sweetening Agents
0
taste receptors, type 1
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
22238Informations de copyright
© 2021. The Author(s).
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