Identification of Opioid-Binding Materials of Rat Brain

概要

論文の詳細を見る
Digitonin-solubilized opioid receptors from rat brain were purified with an affinity resin, AH-Sepharose coupled with [D-Ala^2,D-Ler^5]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 ^<125>I-labeled β-endorphin using bis[2-(succinimidooxycarbonyloxy)-ethyl]sulfone aqs 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 suggested that the purified materials contained opioid-binding materials (opioid receptors).
1990-01-25