The sweet receptor, composed of the monomers T1R2+T1R3, is the principal receptor responsible for sweet taste in mammals. T1R2 and T1R3 are two related G protein-coupled receptors of the T1R type I taste receptor family. Together, T1R2+T1R3 respond to all classes of sweet compounds. T1Rs have a large extracellular domain consisting of a Venus flytrap module and a cysteine-rich domain, a 7-transmembranes domain, and a short C-terminal tail. The primary goal of this application is to elucidate structure-function relationships underlying human T1R2+T1R3 dimeric receptor interactions with the sweet tasting protein monellin. I will take advantage of the fact that monellin acts on the human sweet receptor, but not on the mouse receptor, to identify the specific residues responsible for the human receptor's response to monellin. Our previous work with human/mouse chimeras has shown that the amino-terminal domain (ATD) of human T1R2 (hT1R2) is critical for the response to monellin. We also have shown that the cysteine-rich domain (CRD) of T1R3 plays a role in monellin's activation of the receptor.
The first aim of this application is to identify residues within the ATD of T1R2 critical for sweet receptor sensitivity to monellin. To accomplish this aim 1 will use chimeras of T1R2 containing varying parts of human and mouse T1R2, and carry out site-directed mutagenesis.
The second aim i s to identify residues within the CRD of hT1R3 critical for sweet receptor sensitivity to monellin.
This aim will be accomplished by directed mutagenesis of the CRD based on our initial identification of residues in this region of the receptor that are critical for responses to monellin. Knowledge of the structure/function relationship of T1Rs may be valuable in the development of novel, low caloric sweeteners to help fight against diet-related disorders such as obesity and diabetes. ? ?
| Gupta, Achla; Mulder, Jan; Gomes, Ivone et al. (2010) Increased abundance of opioid receptor heteromers after chronic morphine administration. Sci Signal 3:ra54 |
| Assadi-Porter, Fariba M; Maillet, Emeline L; Radek, James T et al. (2010) Key amino acid residues involved in multi-point binding interactions between brazzein, a sweet protein, and the T1R2-T1R3 human sweet receptor. J Mol Biol 398:584-99 |
| Maillet, Emeline L; Margolskee, Robert F; Mosinger, Bedrich (2009) Phenoxy herbicides and fibrates potently inhibit the human chemosensory receptor subunit T1R3. J Med Chem 52:6931-5 |
| Assadi-Porter, Fariba M; Tonelli, Marco; Maillet, Emeline et al. (2008) Direct NMR detection of the binding of functional ligands to the human sweet receptor, a heterodimeric family 3 GPCR. J Am Chem Soc 130:7212-3 |
| Romanov, Roman A; Rogachevskaja, Olga A; Bystrova, Marina F et al. (2007) Afferent neurotransmission mediated by hemichannels in mammalian taste cells. EMBO J 26:657-67 |
