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Science 5 October 1990:
Vol. 250. no. 4977, pp. 121 - 123
DOI: 10.1126/science.2171146
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Articles
Science, Vol 250, Issue 4977, 121-123
Copyright © 1990 by American Association for the Advancement of Science
Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit
K King,
HG Dohlman,
J Thorner,
MG Caron,
and
RJ Lefkowitz
Department of Medicine (Cardiology), Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710.
To facilitate functional and mechanistic studies of receptor-G protein interactions, [corrected] the human beta 2-adrenergic receptor (h beta-AR) has been expressed in Saccharomyces cerevisiae. This was achieved by placing a modified h beta-AR gene under control of the galactose-inducible GAL1 promoter. After induction by galactose, functional h beta-AR was expressed at a concentration several hundred times as great as that found in any human tissue. As determined from competitive ligand binding experiments, h beta-AR expressed in yeast displayed characteristic affinities, specificity, and stereoselectivity. Partial activation of the yeast pheromone response pathway by beta-adrenergic receptor agonists was achieved in cells coexpressing h beta-AR and a mammalian G protein (Gs) alpha subunit-demonstrating that these components can couple to each other and to downstream effectors when expressed in yeast. This in vivo reconstitution system provides a new approach for examining ligand binding and G protein coupling to cell surface receptors.
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