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Science


Title:Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit
Author(s):King K; Dohlman HG; Thorner J; Caron MG; Lefkowitz RJ;
Address:"Department of Medicine (Cardiology), Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710"
Journal Title:Science
Year:1990
Volume:250
Issue:4977
Page Number:121 - 123
DOI: 10.1126/science.2171146
ISSN/ISBN:0036-8075 (Print) 0036-8075 (Linking)
Abstract:"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"
Keywords:Amino Acid Sequence Base Sequence Cell Membrane/physiology GTP-Binding Proteins/genetics/*physiology Gene Expression Humans Iodocyanopindolol Kinetics Macromolecular Substances Molecular Sequence Data Pindolol/analogs & derivatives/metabolism Plasmids Pro;
Notes:"MedlineKing, K Dohlman, H G Thorner, J Caron, M G Lefkowitz, R J eng GM21841/GM/NIGMS NIH HHS/ HL16037/HL/NHLBI NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. 1990/10/05 Science. 1990 Oct 5; 250(4977):121-3. doi: 10.1126/science.2171146"

 
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