Smart E J, Ying Y S, Conrad P A, Anderson R G
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235.
J Cell Biol. 1994 Dec;127(5):1185-97. doi: 10.1083/jcb.127.5.1185.
Caveolae are a membrane specialization used to internalize molecules by potocytosis. Caveolin, an integral membrane protein, is associated with the striated coat present on the cytoplasmic surface of the caveolae membrane. We now report that oxidation of caveolar cholesterol with cholesterol oxidase rapidly displaces the caveolin from the plasma membrane to intracellular vesicles that colocalize with Golgi apparatus markers. After the enzyme is removed from the medium, caveolin returns to caveolae. When untreated cells are gently homogenized, caveolin on the plasma membrane is accessible to both anti-caveolin IgG and trypsin. After cholesterol oxidase treatment, however, Golgi-associated caveolin is inaccessible to both of these molecules. Brefeldin A, which inhibits ER to Golgi trafficking, blocks the appearance of caveolin in the Golgi apparatus but does not prevent caveolin from leaving the plasma membrane. Indirect immunogold localization experiments show that in the presence of cholesterol oxidase caveolin leaves the plasma membrane and becomes associated with endoplasmic reticulum and Golgi compartments. Surprisingly, the loss of caveolin from the plasma membrane does not affect the number or morphology of the caveolae.
小窝是一种膜特化结构,用于通过胞饮作用内化分子。小窝蛋白是一种整合膜蛋白,与小窝膜细胞质表面的条纹状衣被相关。我们现在报道,用胆固醇氧化酶氧化小窝胆固醇会迅速使小窝蛋白从质膜转移到与高尔基体标记物共定位的细胞内囊泡中。从培养基中去除该酶后,小窝蛋白会回到小窝。当对未处理的细胞进行轻柔匀浆时,质膜上的小窝蛋白可被抗小窝蛋白IgG和胰蛋白酶识别。然而,胆固醇氧化酶处理后,与高尔基体相关的小窝蛋白对这两种分子均不可见。抑制内质网到高尔基体运输的布雷菲德菌素A可阻止小窝蛋白出现在高尔基体中,但不能阻止小窝蛋白离开质膜。间接免疫金定位实验表明,在胆固醇氧化酶存在的情况下,小窝蛋白离开质膜并与内质网和高尔基体区室相关联。令人惊讶的是,质膜上小窝蛋白的缺失并不影响小窝的数量或形态。