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
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 11:05
Last modification date
20/08/2019 12:24