Zukin R S, Kream R M
Proc Natl Acad Sci U S A. 1979 Apr;76(4):1593-7. doi: 10.1073/pnas.76.4.1593.
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.
通过交联从大鼠脑中溶解的非共价复合物，制备了共价结合的[³H]D-丙氨酸²，甲硫氨酸⁵-脑啡肽酰胺和¹²⁵I标记的D-丙氨酸²，N-甲基苯丙氨酸⁴，甲硫氨酸（O）⁵-脑啡肽-大分子复合物。在非变性条件下对部分纯化的¹²⁵I标记的脑啡肽-大分子复合物进行凝胶电泳，得到一个单一的主要/放射性峰。通过分子排阻色谱法测定，该复合物的斯托克斯半径约为48 Å；对于球形分子，该半径对应于380,000的分子量。在初步实验中，该复合物的十二烷基硫酸钠电泳显示出一个主要的放射性峰，对应于35,000的分子量。这些特定的共价脑啡肽-大分子复合物的制备在受体纯化和探索阿片类药物作用的分子机制方面应该是有用的。
