Chemical crosslinking of a solubilized enkephalin macromolecular complex

R. S. Zukin, R. M. Kream

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Abstract

Covalently bound [3H]D-Ala2,Met5-enkephalinamide- and 125I-labeled D-Ala2,N-Me-Phe4,Met-(O)5-ol-enkephalin-macromolecule complexes have been prepared by crosslinking the solubilized noncovalent complexes from rat brain. Gel electrophoresis of the partially purified 125I-labeled enkephalin-macromolecule complex under nondenaturing conditions results in a single major radioactive peak. The complex has a Stokes radius of approximately 48 Å as determined by molecular exclusion chromatography; this radius corresponds to a molecular weight of 380,000 for a spherical molecule. In preliminary experiments, sodium dodecyl sulfate electrophoresis of the complex shows a major radioactive peak corresponding to a molecular weight of 35,000. The preparation of these specific covalent enkephalin-macromolecule complexes should be useful in purification of the receptor and in probing the molecular mechanism of opiate action.

Original languageEnglish (US)
Pages (from-to)1593-1597
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume76
Issue number4
DOIs
StatePublished - Jan 1 1979

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