Bitter tastes are detected at the cellular level by a diverse family of taste receptors (TAS2Rs) belonging to the G protein-coupled receptor (GPCR) superfamily. The lack of direct structural information for TAS2Rs (and most GPCRs) limits our understanding of their structural features responsible for ligand binding and signaling. Using a high-throughput approach called Shotgun Mutagenesis Mapping, we created a comprehensive TAS2R16 mutation library with defined point mutations at every amino acid in the receptor and screened each mutant's functional activity in response to unique ligands, both agonists and inhibitors, using a Ca2+-flux signaling assay. We identified amino acids that, when substituted, abrogate signaling for all agonists, as well as residues important for the activity of only specific ligands. These critical residues define both the binding sites for various TAS2R16 ligands as well as motifs important for signal transduction, and will help guide the structural understanding of bitter taste receptor function.

Comprehensive mapping of functional sites for agonists and inhibitors of the bitter taste receptor TAS2R16 by shotgun mutagenesis

244th National Meeting (2012)

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