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Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris
Author(s) -
Talmont Franck,
Sidobre Stéphane,
Demange Pascal,
Milon Alain,
Emorine Laurent J.
Publication year - 1996
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(96)00971-4
Subject(s) - pichia pastoris , yeast , saccharomyces cerevisiae , opioid receptor , biochemistry , alcohol oxidase , diprenorphine , biology , chemistry , receptor , microbiology and biotechnology , opioid , gene , recombinant dna
The human μ‐opioid receptor cDNA from which the 32 amino‐terminal codons were substituted by the Saccharomyces cerevisiae α‐mating factor signal sequence has been expressed in the methylotrophic yeast Pichia pastoris using the host promoter of the alcohol oxidase‐1 gene. Cell membranes exhibited specific and saturable binding of the opioid antagonist [ 3 H]diprenorphine ( K d = 0.2 nM and B max = 400 fmol/mg protein or 800 sites/cell). Competition studies with non‐selective, and μ‐, δ‐ and κ‐selective opioid agonists and antagonists revealed a typical μ‐opioid receptor binding profile, suggesting proper folding of the protein in yeast membranes.