Publication Date:
1990-10-05
Description:
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.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉King, K -- Dohlman, H G -- Thorner, J -- Caron, M G -- Lefkowitz, R J -- GM21841/GM/NIGMS NIH HHS/ -- HL16037/HL/NHLBI NIH HHS/ -- New York, N.Y. -- Science. 1990 Oct 5;250(4977):121-3.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Medicine (Cardiology), Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/2171146" target="_blank"〉PubMed〈/a〉
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
;
Promoter Regions, Genetic
;
Receptors, Adrenergic, beta/genetics/metabolism/*physiology
;
Recombinant Fusion Proteins/metabolism
;
Restriction Mapping
;
Saccharomyces cerevisiae/genetics/*physiology
;
*Signal Transduction
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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