Open AccessChemical crosslinking of a solubilized enkephalin macromolecular complex.
Author(s) -
R. Suzanne Zukin,
Richard M. Kream
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.4.1593
Subject(s) - macromolecule , stokes radius , chemistry , covalent bond , sodium dodecyl sulfate , enkephalin , size exclusion chromatography , electrophoresis , molecular mass , chromatography , biochemistry , receptor , organic chemistry , enzyme , opioid
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.