title: Introduction of a negative charge at Arg82 in thaumatin abolished responses to human T1R2-T1R3 sweet receptors.
abstract: Thaumatin, an intensely sweet-tasting protein, elicits a sweet-taste sensation at a level as low as 50 nM. Although previous sensory analyses have suggested that Lys67 and Arg82 are important to the sweetness of thaumatin, the exact effects of each residue on sweet receptors are still unknown. In the present study, various mutants of thaumatin altered at Arg82 as well as Lys67 were prepared and their sweetness levels were quantitatively evaluated by cell-based assays using HEK293 cells expressing human sweet receptors. Mutations at Arg82 had a more deteriorative effect on sweetness than mutations at Lys67. Particularly, a charge inversion at Arg82 (R82E) resulted in an abolishment of the response to sweet receptors even at a concentration as high as 1mM. These results indicate that Arg82 plays a central role in determining the sweetness of thaumatin. A strict spatial charge location at residue 82 appears to be required for interaction with sweet receptors.
authors.affiliation: Department of Natural Resources, Graduate School of Global and Environmental Studies, Kyoto University, Uji, Kyoto 611-0011, Japan. k_ooota@kais.kyoto-u.ac.jp
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ohta
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Masuda
authors.forename: Fumito
authors.initials: F
authors.lastname: Tani
authors.forename: Naofumi
authors.initials: N
authors.lastname: Kitabatake
mesh_terms: D001120:Arginine / Q000737:chemistry / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D010843:Pichia; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D013549:Sweetening Agents / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D043562:Receptors, G-Protein-Coupled; D013549:Sweetening Agents; C431926:taste receptors, type 1; C003427:thaumatin protein, plant; D001120:Arginine