Hitomi K, Masuda S, Ito K, Ueda M, Sasaki R
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
Biochem Biophys Res Commun. 1989 May 15;160(3):1140-8. doi: 10.1016/s0006-291x(89)80122-6.
Functional erythropoietin (Ep) receptor (Ep-R) on transplantable mouse leukemic cells was solubilized by 3[(3-cholamidopropyl)dimethyl ammonio]-1-propanesulfate (CHAPS). An assay of the solubilized Ep-R was established: (1) radioiodinated Ep was bound to the solubilized Ep-R, (2) Ep.Ep-R complexes were covalently cross-linked, (3) the cross-linked products were selectively precipitated by polyethylene glycol, (4) the products were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and (5) the Ep.Ep-R cross-linked products were measured. Optimal conditions to yield the cross-linked products and to stabilize the solubilized Ep-R were found. The solubilized Ep-R had properties similar to those of Ep-R on the membrane with respect to binding with Ep and the cross-linked products. These results would be exploited to allow purification of Ep-R.
可移植性小鼠白血病细胞上的功能性促红细胞生成素（Ep）受体（Ep-R）用3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐（CHAPS）进行增溶。建立了一种对增溶后的Ep-R的检测方法：（1）将放射性碘化的Ep与增溶后的Ep-R结合，（2）将Ep-Ep-R复合物进行共价交联，（3）用聚乙二醇选择性沉淀交联产物，（4）在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上分离产物，（5）测定Ep-Ep-R交联产物。发现了产生交联产物和稳定增溶后的Ep-R的最佳条件。增溶后的Ep-R在与Ep结合及交联产物方面具有与膜上Ep-R相似的特性。这些结果将用于Ep-R的纯化。
