Production of adrenergic receptors in yeast

Dorothea Sizmann, Arja Kuusinen, Sirkka Keränen, J. Lomasney, M. Caron

Research output: Contribution to journalArticleScientificpeer-review

24 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)197-203
JournalReceptors and Channels
Volume4
Issue number3
Publication statusPublished - 1996
MoE publication typeA1 Journal article-refereed

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