The human β₂‐adrenergic receptor expressed in Schizosaccharomyces pombe retains its pharmacological properties

Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris

“…The receptor densities determined in this work are somewhat lower than those reported for other G-protein-coupled receptors expressed in various species of yeast [9,11,13,15]. Several studies have documented the utility of signal sequences for the expression of such receptors in yeast.…”

“…In similar experiments performed on whole cells the Kd was 0.17+0.02 nM and the receptor density 810 sites/cell (not shown). Such receptor densities are in the same range as those obtained for expression of Ml-and M5-muscarinic receptors [10,12] in yeast but lower than for 132-adrenergic, D2-dopaminergic and 5HTr, A-serotoninergic receptors [11,[13][14][15]. Utilization of protease-deficient strains has been suggested to increase expression levels of G-protein-coupled receptors in S. cerevisiae [11] and P. pastoris [9].…”

“…In an initial study it was shown that with no modifications of its amino-terminus, the human Ml-muscarinic receptor was poorly expressed in S. cerevisiae (20 fmol/mg protein) [10]. Subsequently, it was suggested that higher expression levels could be reached by replacing the amino-terminal part of the receptors by the corresponding region of the STE2 gene [11,13,15]. Since then, other amino-terminal fusion partners, such as the signal sequences for the c~-mating factor [9,12] or the acid phosphatase [9] have been successfully used.…”

“…Moreover, despite the cloning of numerous genes for G-protein-coupled receptors, only five of these have been expressed in yeast [9][10][11][12][13][14][15]. In this paper, we report on the expression of the human g-opioid receptor (HUMOR) in the methylotrophic yeast P. pastoris.…”

Heterologous expression of G-protein-coupled receptors: comparison of expression systems from the standpoint of large-scale production and purification

“…pombe [53]. However, the neurokinin NK2 receptor was only produced at a level of 1.2 pmol per milligram of membrane protein [54], and the b 2 -adrenergic at 7.5 pmol per milligram of membrane protein [55]. In the case of the NK2 receptor, the fusion sequence used was that of the signal sequence of the acid phosphatase PhoI, a yeast protein located at the cell surface.…”