RT Journal Article
SR Electronic
T1 Affinity labeling of mu opioid receptors by sulfhydryl alkylating derivatives of morphine and morphinone.
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP 50
OP 59
VO 37
IS 1
A1 J M Bidlack
A1 D K Frey
A1 R A Kaplan
A1 A Seyed-Mozaffari
A1 S Archer
YR 1990
UL http://molpharm.aspetjournals.org/content/37/1/50.abstract
AB After reduction of a disulfide bond at or near the mu opioid binding site in rat brain membranes, incubating membranes with 14 beta-bromoacetamido derivatives of either morphine, dihydromorphine, morphinone, or dihydromorphinone resulted in the irreversible inhibition of mu opioid binding to rat brain membranes. Without the addition of the disulfide bond-reducing reagent dithiothreitol, these affinity ligands bound reversibly to opioid binding sites. Binding to either delta or kappa opioid binding sites was not altered by alkylation of the membranes with the affinity ligands. The percentage of irreversible inhibition of mu opioid binding was dependent on the time and temperature of the incubation of membranes with the affinity ligands and on the concentrations of dithiothreitol and the affinity ligands. Incubating membranes with morphine afforded almost complete protection from alkylation of the mu opioid binding site. Naloxone and the l-isomer levorphanol also protected the site from alkylation, whereas the d-isomer dextrorphan and the kappa-selective opioid U50,488H did not protect the site. The mu-selective peptide [D-Ala2, (Me)Phe4,Gly(ol)5]enkephalin was the peptide that afforded the greatest protection. These studies have shown that, after the reduction of a disulfide bond at or near the mu opioid binding site, this sulfhydryl group can be specifically alkylated, resulting in the affinity labeling of the mu opioid binding site.