Method of identifying bitter compounds by employing TRP8, a transient receptor potential channel expressed in taste receptor cells
View Patent ↗The present invention relates to the discovery, identification and characterization of a transient receptor potential channel, referred to herein as TRP8, which is expressed in taste receptor cells and associated with the perception of bitter and sweet taste. The invention encompasses TRP8 nucleotides, host cell expression systems, TRP8 proteins, fusion proteins, polypeptides and peptides, antibodies to the TRP8 protein, transgenic animals that express a TRP8 transgene, and recombinant “knock-out” animals that do not express TRP8. The invention further relates to methods for identifying modulators of the TRP8-mediated taste response and the use of such modulators to either inhibit or promote the perception of bitterness or sweetness. The modulators of TRP8 activity may be used as flavor enhancers in foods, beverages and pharmaceuticals.
1. A method for identifying a compound that potentially induces the perception of a bitter taste comprising:
(i) contacting an isolated cell expressing the TRP8 channel protein of SEQ ID NO: 4 with a test compound and measuring the level of TRP8 activation;
(ii) in a separate experiment, contacting an isolated cell expressing the TRP8 channel protein of SEQ ID NO: 4 with a vehicle control and measuring the level of TRP8 activation where the conditions are essentially the same as in part (i); and
(iii) comparing the level of activation of TRP8 measured in part (i) with the level of activation of TRP8 in part (ii),
wherein the level of TRP8 activation is measured by measuring the membrane potential of the cell and wherein an increased level of activated TRP8 in the presence of the test compound indicates that the test compound potentially induces the perception of a bitter taste.
2. The method according to claim 1 , wherein said measuring the level of TRP8 activation is carried out with one or more fluorescence-indicator dyes.
3. The method according to claim 1 , wherein said measuring the membrane potential of the cell is carried out under voltage clamp assay conditions.
4. The method according to claim 1 , wherein said measuring the membrane potential of the cell is carried out under patch recording assay conditions.