Chemical crosslinking of a solubilized enkephalin macromolecular complex.
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 A 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.
D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism; D004120:Dimethyl Suberimidate; D004723:Endorphins / Q000378:metabolism; D004745:Enkephalins / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D011957:Receptors, Opioid / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism
D004723:Endorphins; D004745:Enkephalins; D011957:Receptors, Opioid; D004120:Dimethyl Suberimidate