Radioiodination and binding characteristics of a highly active opioid peptide

R. M. Kream, R. S. Zukin

Research output: Contribution to journalArticlepeer-review


D-ala2, N-Me-phe4, met (O)5-ol-enkephalin (Sandoz, FK 33-824) was radiolabeled to a specific activity of 1000 Ci/mmol using soluble lactoperoxidase and carrier-free Na125l. The [125l]-labeled enkephalin analog was purified by a combination of gel filtration and ion-exchange chromatography; it cochromatographed with its [127l] labeled analog on thin layers. [125l] D-ala2, N-Me-phe4, met (O)5-ol-enkephalin binds stereospecifically to whole brain membrane preparations from rat with saturation at 5 nM at 25°C. Scatchard analysis reveals two classes of binding sites with apparent Kd's of 0.42 x 10-9 M and 3.70 x 10-9 M. The ratio of high affinity to low affinity binding sites is approximately 1:3. At 25°C, 40 mM sodium ion elicits half-maximal inhibition of stereospecifically bound counts, while 100 mM potassium inhibits binding by 25%. [125l] FK 33-824 has been used in studies involving the purification of the mammalian opiate receptor. Preliminary characterization of a chemically cross-linked, solubilized [125l] FK 33-824 enkephalin macromolecular complex has been carried out. The native complex has an apparent molecular weight of 380,000; SDS gel electrophoresis of the enkephalin-labeled receptor reveals a major labeled subunit of 35,000 daltons.

Original languageEnglish (US)
Pages (from-to)No.137
JournalFederation Proceedings
Issue number3 I
StatePublished - Jan 1 1979

ASJC Scopus subject areas

  • Medicine(all)


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