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PMID: 2845411 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human beta 2-adrenergic receptors expressed in Escherichia coli membranes retain their pharmacological properties.

Marullo S, Delavier-Klutchko C, Eshdat Y, Strosberg AD, Emorine L

Abstract

The coding region of the gene for the human beta 2-adrenergic receptor gene was fused to the beta-galactosidase gene of the lambda gt11 expression vector. The Y1089 Escherichia coli strain was lysogenized with this modified vector and transcription of the fusion gene was induced. Expression of this transcription unit was shown by the appearance in the bacteria of proteins of molecular weight higher than that of native beta-galactosidase, which are immunoreactive with anti-beta-galactosidase antibodies. Production of beta 2-adrenergic receptors was shown by the presence, on intact bacteria, of binding sites for catecholamine agonists and antagonists possessing a typical beta 2-adrenergic pharmacological profile. Binding and photoaffinity labeling studies performed on intact E. coli and its membrane fractions showed that these binding sites are located in the inner membrane of the bacteria. Expression of pharmacologically active human beta 2-adrenergic receptors in E. coli further supports the similar transmembrane organization proposed for bacteriorhodopsin and eukaryotic membrane-embedded receptors coupled to guanine nucleotide-binding regulatory proteins. Moreover, this system should facilitate future analyses of the ligand-binding properties within this family of membrane receptors.

MeSH Terms
Affinity Labels Autoradiography Cell Fractionation Cell Membrane/metabolism Centrifugation, Density Gradient Cloning, Molecular Escherichia coli/genetics,metabolism,ultrastructure Gene Expression Regulation Genetic Vectors Humans Immunoblotting Ligands Receptors, Adrenergic, beta/biosynthesis,genetics,metabolism Transcription, Genetic beta-Galactosidase/genetics
Chemicals
Affinity Labels Ligands Receptors, Adrenergic, beta beta-Galactosidase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Marullo S
Centre National de la Recherche Scientifique, Université Paris VII, France.
Delavier-Klutchko C
Eshdat Y
Strosberg A D
Emorine L
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1988-10-00
Pages
7551-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC282229
Subset
IM
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