科研之友
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摘要
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 -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 -arrestin-mediated endosomal receptor internalization can be monitored by ligand-induced enzyme complementation of -galactosidase (-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 -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.
- 出版日期2018-3
