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

Détails

ID Serval
serval:BIB_0202D9FA5B23
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Expression of the alpha 1b-adrenergic receptor and G protein subunits in mammalian cell lines using the Semliki Forest virus expression system.
Périodique
Journal of Receptor and Signal Transduction Research
Auteur⸱e⸱s
Scheer A., Björklöf K., Cotecchia S., Lundstrom K.
ISSN
1079-9893 (Print)
ISSN-L
1079-9893
Statut éditorial
Publié
Date de publication
1999
Volume
19
Numéro
1-4
Pages
369-378
Langue
anglais
Résumé
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.
Mots-clé
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
Création de la notice
24/01/2008 12:05
Dernière modification de la notice
20/08/2019 13:24
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