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

J Recept Signal Transduct Res. 1999 Jan-Jul;19(1-4):369-78. doi: 10.3109/10799899909036658.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • COS Cells
  • Cell Line
  • Cricetinae
  • GTP-Binding Proteins / biosynthesis
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / genetics*
  • Gene Expression
  • Genetic Vectors
  • Inositol Phosphates / metabolism
  • Kinetics
  • Protein Conformation
  • Receptors, Adrenergic, alpha-1 / biosynthesis
  • Receptors, Adrenergic, alpha-1 / chemistry*
  • Receptors, Adrenergic, alpha-1 / genetics*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Semliki forest virus / genetics

Substances

  • Inositol Phosphates
  • Receptors, Adrenergic, alpha-1
  • Recombinant Proteins
  • GTP-Binding Proteins