RAPID PARTIAL-PURIFICATION OF PLACENTAL GLUCOCEREBROSIDE BETA-GLUCOSIDASE AND ITS ENTRAPMENT IN LIPOSOMES
RAPID PARTIAL-PURIFICATION OF PLACENTAL GLUCOCEREBROSIDE BETA-GLUCOSIDASE AND ITS ENTRAPMENT IN LIPOSOMES
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DOI：
10.1042/bj1640439
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发表时间：
1977-01-01
影响因子：
4.1
通讯作者：
GREGORIADIS, G
中科院分区：
文献类型：
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作者：
BRAIDMAN, IP;GREGORIADIS, G
葡糖脑苷脂β-本文采用刀豆球蛋白A-Sepharose亲和层析和有机溶剂沉淀相结合的方法，从人胎盘中快速获得富含葡萄糖醛酸酶的无洗涤剂制剂。在典型的制备中，在2天内从5个胎盘获得约11，000单位的纯化1500倍的酶。该酶制剂还含有其他水解酶，但它们的纯化程度要小得多。对脂质体中包封的研究表明，所有葡糖脑苷脂β-将所用的葡糖苷酶活性掺入中性卵磷脂酰胆碱-胆固醇脂质体中。与脂质体的协会歧视其他蛋白质，包括一些水解酶，从而有助于进一步纯化的酶。超过95%的脂质体相关酶活性是潜在的。
A glucocerebroside .beta.-glucosidase-rich detergent-free preparation was obtained from human placentas by a rapid method combining affinity chromatography on concanavalin A-Sepharose and organic-solvent precipitation. In a typical preparation about 11,000 units of the enzyme purified 1500-fold were obtained from 5 placentas in 2 days. The enzyme preparation also contained other hydrolases, but the extent of their purification was much smaller. Studies on entrapment in liposomes showed that all glucocerebroside .beta.-glucosidase activity used was incorporated in neutral egg phosphatidylcholine-cholesterol liposomes. Association with liposomes discriminated against other proteins, including some of the hydrolases, thus contributing to further purification of the enzyme. More than 95% of the liposome-associated enzyme activity was latent.