The human β 2‐adrenergic receptor expressed in Schizosaccharomyces pombe retains its pharmacological properties

FEBS Letters

Testa, Luana¹  Tocchini Valentini, Glauco P.²  Ficca, Anna Grazia¹ 

[1] EniChem SpA, Istituto Guido Donegani, Via E.Ramarini 32, 00016 Monterotondo/Rome, Italy;Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, Viale K Marx 43, 00137 Rome, Italy

关键词: Human β 2-adrenergic receptor;    Yeast vector;    Schizosaccharomyces pombe;    Membrane protein;    Heterologous expression;   

DOI  :  10.1016/0014-5793(95)01330-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have developed a rapid and efficient expression system to study the human β ₂ adrenergic receptor (huβ ₂AR) in the fission yeast Schizosaccharomyces pombe. This was achieved by cloning the huβ ₂AR 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). Huβ ₂AR 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 [¹²⁵I]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 huβ ₂AR. This system constitutes a convenient alternative to existing methods for studying seven transmembrane domain receptors due to its simplicity and high reproducibility.

【 授权许可】

Unknown