Production of adrenergic receptors in yeast.

Published

Journal Article

Using a recombinant yeast strain expressing human beta 2 adrenergic receptors under a galactose-inducible promoter, we established conditions for receptor production in 1-15 liter fermenter culture. Crucial factors contributing to consistent high-level expression included the use of selective glucose-free medium, the maintenance of the pH of the culture at 7.2-7.5 and the presence of an antagonist. The expression strategy and production conditions used with the beta 2 adrenergic receptor were then employed to express the human alpha 2-C2 adrenergic receptor in Saccharomyces cerevisiae. Galactose-induced yeast cells displayed specific, high-affinity [3H]rauwolscine binding and contained a 50-kDa species recognized by an alpha 2-C2 receptor specific antiserum. In fermenter culture, up to 10(5) high-affinity [3H]rauwolscine binding sites per cell (corresponding to 30-60 pmol/mg of protein) were obtained. The high expression level combined with relative ease and low cost of scaling-up make yeast a promising alternative to mammalian cells for the production of adrenergic and other G-protein-coupled receptors for structural studies.

Full Text

Duke Authors

Cited Authors

  • Sizmann, D; Kuusinen, H; Keränen, S; Lomasney, J; Caron, MG; Lefkowitz, RJ; Keinänen, K

Published Date

  • 1996

Published In

Volume / Issue

  • 4 / 3

Start / End Page

  • 197 - 203

PubMed ID

  • 9014242

Pubmed Central ID

  • 9014242

International Standard Serial Number (ISSN)

  • 1060-6823

Language

  • eng

Conference Location

  • England