Expression of the alpha 1b-adrenergic receptor and G protein subunits in mammalian cell lines using the Semliki Forest virus expression system.

Details

Serval ID
serval:BIB_0202D9FA5B23
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Expression of the alpha 1b-adrenergic receptor and G protein subunits in mammalian cell lines using the Semliki Forest virus expression system.
Journal
Journal of Receptor and Signal Transduction Research
Author(s)
Scheer A., Björklöf K., Cotecchia S., Lundstrom K.
ISSN
1079-9893 (Print)
ISSN-L
1079-9893
Publication state
Published
Issued date
1999
Volume
19
Number
1-4
Pages
369-378
Language
english
Abstract
Semliki Forest virus (SFV) vectors have been efficiently used for rapid high level expression of several G protein-coupled receptors. Here we describe the use of SFV vectors to express the alpha 1b-adrenergic receptor (AR) alone or in the presence of the G protein alpha q and/or beta 2 and gamma 2 subunits. Infection of baby hamster kidney (BHK) cells with recombinant SFV-alpha 1b-AR particles resulted in high specific binding activity of the alpha 1b-AR (24 pmol receptor/mg protein). Time-course studies indicated that the highest level of receptor expression was obtained 30 hours post-infection. The stimulation of BHK cells, with epinephrine led to a 5-fold increase in inositol phosphate (IP) accumulation, confirming the functional coupling of the receptor to G protein-mediated activation of phospholipase C. The SFV expression system represents a rapid and reproducible system to study the pharmacological properties and interactions of G protein coupled receptors and of G protein subunits.
Keywords
Animals, CHO Cells, COS Cells, Cell Line, Cricetinae, GTP-Binding Proteins/biosynthesis, GTP-Binding Proteins/chemistry, Gene Expression, Genetic Vectors, Inositol Phosphates/metabolism, Kinetics, Protein Conformation, Receptors, Adrenergic, alpha-1/biosynthesis, Receptors, Adrenergic, alpha-1/chemistry, Recombinant Proteins/biosynthesis, Recombinant Proteins/chemistry, Semliki forest virus/genetics
Pubmed
Web of science
Create date
24/01/2008 12:05
Last modification date
20/08/2019 13:24
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