ACS DIVISION OF BIOLOGICAL CHEMISTRY

July 23rd, 2014 by Sanematsu, K., Kusakabe, Y., Shigemura, N., Hirokawa, T., Nakamura, S., Imoto, T., Ninomiya, Y.

Gymnemic acids are triterpene glycosides that selectively suppress taste responses to various sweet substances in humans, but not in mice. This sweet-suppressing effect of gymnemic acids is diminished by rinsing the tongue with γ-cyclodextrin (γ-CD). However, little is known about the molecular mechanisms underlying sweet-suppressing effect of gymnemic acids and interaction between gymnemic acids vs. sweet taste receptor and/or γ-CD. To investigate whether gymnemic acids directly interact with human sweet receptor hT1R2 + hT1R3, we used the sweet receptor T1R2+T1R3 assay in transiently transfected HEK293 cells. Similar to previous studies in humans and mice, gymnemic acids (100 ug/ml) inhibited the [Ca2+]i responses to sweet compounds in HEK293 cells heterologously expressing hT1R2+hT1R3 but not in those expressing mouse sweet receptor mT1R2 + mT1R3. The effect of gymnemic acids rapidly disappeared after rinsing the HEK293 cells with γ-CD. Using mixed-species pairings of human and mouse sweet receptor subunits and chimeras, we determined that the transmembrane domain of hT1R3 was mainly required for the sweet-suppressing effect of gymnemic acids. Directed mutagenesis in the transmembrane domain of hT1R3 revealed that the interaction site for gymnemic acids shared the amino acid residues which determined the sensitivity to another sweet antagonist, lactisole. Glucuronic acid which is the common structure of gymnemic acids also reduced sensitivity to sweet compounds. In our models, gymnemic acids were predicted to dock to a binding pocket within the transmembrane domain of hT1R3.