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The human β 2 ‐adrenergic receptor expressed in Schizosaccharomyces pombe retains its pharmacological properties
Author(s) -
Ficca Anna Grazia,
Testa Luana,
Tocchini Valentini Glauco P.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01330-x
Subject(s) - schizosaccharomyces pombe , yeast , biology , fusion protein , saccharomyces cerevisiae , receptor , microbiology and biotechnology , gene , biochemistry , recombinant dna
We have developed a rapid and efficient expression system to study the human β 2 adrenergic receptor (hu β 2 AR) in the fission yeast Schizosaccharomyces pombe . This was achieved by cloning the hu β 2 AR gene, modified by replacement of the 5′ untranslated and a small part of the N‐terminal coding sequence (first 14 amino acids) with the corresponding region of the yeast Saccharomyces cerevisiae STE2 (α‐factor receptor) gene. The gene was then placed under the control of a S. pombe constitutive promoter for alcohol dehydrogenase (adh). Hu β 2 AR expression was assessed by immunoblot analysis of the chimeric protein with an anti‐STE2 serum raised against a dodecapeptide homologous to the N‐terminal amino acids of STE2 and ligand binding was assayed using [ 125 I]cyanopindolol. We demonstrate here that the chimeric receptor expressed in S. pombe exhibits the same characteristic ligand specificity and affinity as that of the authentic hu β 2 AR. This system constitutes a convenient alternative to existing methods for studying seven transmembrane domain receptors due to its simplicity and high reproducibility.