A DETERGENT-FREE METHOD FOR PURIFYING CAVEOLAE MEMBRANE FROM TISSUE-CULTURE CELLS
A DETERGENT-FREE METHOD FOR PURIFYING CAVEOLAE MEMBRANE FROM TISSUE-CULTURE CELLS
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DOI：
10.1073/pnas.92.22.10104
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发表时间：
1995-10-24
影响因子：
11.1
通讯作者：
ANDERSON, RGW
中科院分区：
文献类型：
--
作者：
SMART, EJ;YING, YS;ANDERSON, RGW
目前用于从组织培养细胞中纯化小窝的方法利用了该膜结构域的Triton X-100不溶性。为了避免使用清洁剂，我们开发了一种取决于小窝膜独特浮力密度的方法。我们获得的小窝组分高度富集小窝蛋白。因此，我们能够识别以前未被检测到的小窝相关蛋白。此外，可溶于Triton X-100的固有窖蛋白在分离过程中被保留。
Current methods for purifying caveolae from tissue culture cells take advantage of the Triton X-100 insolubility of this membrane domain. To circumvent the use of detergents, we have developed a method that depends upon the unique buoyant density of caveolae membrane. The caveolae fractions that we obtain are highly enriched in caveolin. As a consequence we are able to identify caveolae-associated proteins that had previously gone undetected. Moreover, resident caveolae proteins that are soluble in Triton X-100 are retained during the isolation.