THE HUMAN BETA(2)-ADRENERGIC RECEPTOR EXPRESSED IN SCHIZOSACCHAROMYCES-POMBE RETAINS ITS PHARMACOLOGICAL PROPERTIES

We have developed a rapid and efficient expression system to study the human beta(2) adrenergic receptor (hu beta(2)AR) in the fission yeast Schizosaccharomyces pombe, This was achieved by cloning the hup,AR ge ne, modified by replacement of the 5' untranslated and a small part of the N-terminal coding sequence (first 14 amino acids) with the corres ponding region of the yeast Saccharomyces cerevisiae STE2 (alpha-facto r receptor) gene, The gene was then placed under the control of a S. p ombe constitutive promoter for alcohol dehydrogenase (adh), Hu beta(2) AR expression was assessed by immunoblot analysis of the chimeric prot ein with an anti-STE(2) serum raised against a dodecapeptide homologou s to the N-terminal amino acids of STE2 and ligand binding was assayed using [I-125]cyanopindolol. We demonstrate here that the chimeric rec eptor expressed in S. pombe exhibits the same characteristic ligand sp ecificity and affinity as that of the authentic hu beta(2)AR. This sys tem constitutes a convenient alternative to existing methods for study ing seven transmembrane domain receptors due to its simplicity and hig h reproducibility.