Rapid, high-yield purification of cell surface membrane using colloidal magnetite coated with polyvinylamine: sedimentation versus magnetic isolation.
Rapid, high-yield purification of cell surface membrane using colloidal magnetite coated with polyvinylamine: sedimentation versus magnetic isolation.
复制标题
使用涂有聚乙烯胺的胶体磁铁矿快速、高产地纯化细胞表面膜：沉降与磁力分离。
DOI：
10.1016/0005-2736(85)90396-7
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Jacobson,BS
中科院分区：
文献类型：
--
作者：
Patton,WF;Kim,J;Jacobson,BS
本文介绍了一种新的磁分离Dictyosteelium discoideum外质膜的技术，并与以前发表的采用二氧化硅包被的质膜沉降的方法进行了比较。磁隔离技术包括用聚乙烯胺-磁铁矿胶体涂覆完整的细胞，并用聚丙烯酸酯覆盖以形成致密的薄膜。磁铁矿膜完全覆盖细胞，没有内化。裂解包被的细胞，并使用发散场电磁体从细胞匀浆中回收膜碎片。分析以这种方式获得的膜的标记酶活性和细胞表面标记。原生质膜以高收率（42%）获得，平均纯化8倍。聚乙烯胺-磁铁矿膜屏蔽外部质膜面对木瓜蛋白酶和链霉蛋白酶的蛋白水解。在伴刀豆球蛋白A-碳水化合物竞争研究中，它也作为α-甲基甘露糖苷的屏障。
A new technique for the magnetic isolation of external plasma membrane fromDictyostelium discoideumis described and compared to a previously published procedure employing sedimentation of silica-coated plasma membrane. The magnetic isolation technique involves coating intact cells with a polyvinylamine-magnetite colloid and overcoating with polyacrylate to form a dense pellicle. The magnetite pellicle totally coated the cells and was not internalized. Coated cells were lysed and membrane fragments retrieved from the cell homogenate using a diverging field electromagnet. The membrane obtained in such a manner was analyzed for marker enzyme activity and cell surface label. The plasma membrane was obtained in high yield (42%) with an average purification of 8-fold. The polyvinylamine-magnetite pellicle shielded the external plasma membrane face to proteolysis by papain and pronase. It also acted as a barrier to α-methylmannoside in concanavalin A-carbohydrate competition studies.