MOLECULAR CLONING OF THE HUMAN SWEET TASTE RECEPTOR

Project: Research project

Project Details

Description

The sense of taste provides animals with a rapid, though limited chemical
analysis of a potential food substance. Based on this information the
animal decides whether to ingest or expel the substance. In humans,
disorders of taste lead to anorexia and subsequent weight loss, which
complicates the management of the underlying diseases. The pathophysiology
of taste disorders is poorly understood because little is known about the
molecular mechanisms involved in the transduction of any taste modality.
Our previous studies have demonstrated that bitter taste transduction
involves cell surface receptors. Binding of ligand to these receptors
stimulates release of calcium from internal stores suggesting the
involvement of the inositol triphosphate pathway in the bitter transduction
process. The transduction of sweet taste appears to involve a different
mechanism. Purification or molecular cloning of the taste receptors would
enhance our understanding of the primary events in taste transduction,
however, to date none of the taste receptors have been purified or cloned.

The long-term goal of this investigation is to elucidate the molecular
mechanisms involved in sweet taste transduction. To accomplish this goal,
the technique of DNA-mediated gene transfer will be used to establish a
mouse cell line expressing the human sweet taste receptor. To assay for
expression of the human sweet taste receptor the two sweet proteins,
thaumatin and monellin, will be used in an erythrocyte rosetting assay
utilizing anti- thaumatin antibodies coupled to the erythrocytes to detect
transformed mouse L-cells expressing the human thaumatin-binding sweet
taste receptor. Following establishment of the cell line, the gene for the
sweet taste receptor will be molecularly cloned from this cell line. The
cell line will also facilitate studies of the mechanism of sweet taste
transduction to identify second messenger processes involved in the
transduction process.

The specific goals of this proposal are: 1) To establish a mouse L-cell
line expressing the human sweet taste receptor by the technique of DNA-
mediated gene transfer; 2) To utilize the cell line to molecularly clone
the human sweet taste receptor, and 3) To utilize the cell line to study
the mechanism of sweet taste transduction.
StatusFinished
Effective start/end date1/1/9212/31/92

Funding

  • National Institute on Deafness and Other Communication Disorders

ASJC

  • Cellular and Molecular Neuroscience
  • Molecular Medicine
  • Biophysics
  • Endocrinology, Diabetes and Metabolism
  • Cell Biology

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