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" |
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" |