Chem. Senses 25: 361-368,
2000
© Oxford University Press 2000
An In Vitro Assay Useful to Determine the Potency of Several Bitter Compounds
Howard Hughes Medical Institute, Department of Physiology and Biophysics, The Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029, USA 1 Present address: Almirall Prodesfarma Cardener 64, E-08022 Barcelona, Spain 2 Present address: Global Nutrition R & D, Novartis Consumer Health Company, 560 Morris Avenue, Summit, NJ 07901-1312, USA
Correspondence to be sent to: Robert F. Margolskee, Howard Hughes Medical Institute, Department of Physiology and Biophysics, The Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029, USA. e-mail bob{at}inka.mssm.edu
Gustducin and transducin are guanine nucleotide binding regulatory proteins (G proteins) expressed in taste receptor cells and implicated in transducing taste cell responses to certain compounds that humans consider bitter or sweet. These G proteins can be activated in vitro by taste receptor-containing membranes plus any of several bitter compounds. This activation can be monitored using limited trypsin digestion, sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting. Scanning of the autoradiograms enables one to quantitate the level of activation (defined as an activation index), obtain dose–response profiles and estimate the potency of the tastant. This assay may provide a useful substitute for, or adjunct to, the time-consuming human psychophysical analysis and costly animal studies typically used in taste sensory analysis. It may be used to identify and determine the concentration–response function of many bitter components of oral pharmaceuticals and food ingredients. A potential limitation of the assay is that only about half of all bitter compounds tested demonstrated in vitro activity, perhaps due to the presence of multiple transduction pathways. Nevertheless, the rapid throughput and microsample handling capability of this assay make it an ideal method to screen for high-potency bitterness inhibitors.
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