title: Identification of opioid-binding materials of rat brain.
abstract: Digitonin-solubilized opioid receptors from rat brain were purified with an affinity resin, AH-Sepharose coupled with [D-Ala2, D-Leu5]enkephalin (DADLE). Radioreceptor binding assay showed that the purified materials had specific opioid-binding activity of 310 pmol/mg protein on DADLE binding. Analyses by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) revealed that the materials were rich in two polypeptides; the major component had a molecular weight of 62000-64000. To establish the materials responsible for binding opiates, the purified materials were cross-linked with 125I-labeled beta-endorphin using bis[2-(succinimidooxycarbonyloxy)-ethyl]sulfone as a cross-linker. The molecular weight of 62000-64000, the major band of the purified materials on SDS-PAGE, agreed closely with that determined by the cross-linking experiment. The results suggest that the purified materials contained opioid-binding materials (opioid receptors).
authors.affiliation: Faculty of Pharmaceutical Sciences, Mukogawa Women's University, Hyogo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Fujioka
authors.forename: F
authors.initials: F
authors.lastname: Inoue
authors.forename: M
authors.initials: M
authors.lastname: Kuriyama
authors.forename: Y
authors.initials: Y
authors.lastname: Miura
authors.forename: T
authors.initials: T
authors.lastname: Higashijima
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D004743:Enkephalin, Leucine / Q000031:analogs & derivatives / Q000378:metabolism; D016308:Enkephalin, Leucine-2-Alanine; D051381:Rats; D011957:Receptors, Opioid / Q000032:analysis / Q000302:isolation & purification*
chemical_list: D011957:Receptors, Opioid; D004743:Enkephalin, Leucine; D016308:Enkephalin, Leucine-2-Alanine