Kinoshita T, Nachman R L, Minick R
J Cell Biol. 1979 Sep;82(3):688-96. doi: 10.1083/jcb.82.3.688.
Human platelet plasma membranes were isolated with polylysine beads according to the technique developed by Jacobson and Branton (1977, Science [Wash. D. C.] 195:302--304). Lactoperoxidase-catalyzed surface iodination revealed that ninefold greater 125I specific activity was associated with the membranes isolated on beads than with whole platelets. Enrichment in the bead membrane preparation of the activities of membrane marker enzymes, bis(p-nitrophenyl)phosphate phosphodiesterase and Na,K-ATPase, was 8.0 and 4.4, respectively. Contamination with enzymes of other organelles, cytochrome oxidase and beta-glucuronidase, was relatively low as compared with membranes isolated by sucrose gradient centrifugation. Analysis by SDS polyacrylamide gel electrophoresis showed that a full complement of surface glycoproteins was present on the membranes isolated with polylysine beads. The polylysine bead technique is a rapid, reproducible and efficient method for the preparation of relatively pure platelet plasma membranes.
根据雅各布森和布兰顿(1977年,《科学》[华盛顿特区]195:302 - 304)开发的技术,用聚赖氨酸珠分离人血小板质膜。乳过氧化物酶催化的表面碘化显示,与珠上分离的膜相关的125I比活比全血小板高九倍。膜标记酶双(对硝基苯基)磷酸磷酸二酯酶和钠钾ATP酶在珠膜制备物中的富集倍数分别为8.0和4.4。与通过蔗糖梯度离心分离的膜相比,其他细胞器的酶细胞色素氧化酶和β-葡萄糖醛酸酶的污染相对较低。十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析表明,用聚赖氨酸珠分离的膜上存在完整的表面糖蛋白。聚赖氨酸珠技术是一种制备相对纯的血小板质膜的快速、可重复且高效的方法。