Luetterforst R, Stang E, Zorzi N, Carozzi A, Way M, Parton R G
Centre for Microscopy and Microanalysis, Department of Physiology and Pharmacology, and Centre for Molecular and Cellular Biology, University of Queensland, Brisbane, Queensland 4072, Australia.
J Cell Biol. 1999 Jun 28;145(7):1443-59. doi: 10.1083/jcb.145.7.1443.
Caveolins are integral membrane proteins which are a major component of caveolae. In addition, caveolins have been proposed to cycle between intracellular compartments and the cell surface but the exact trafficking route and targeting information in the caveolin molecule have not been defined. We show that antibodies against the caveolin scaffolding domain or against the COOH terminus of caveolin-1 show a striking specificity for the Golgi pool of caveolin and do not recognize surface caveolin by immunofluorescence. To analyze the Golgi targeting of caveolin in more detail, caveolin mutants were expressed in fibroblasts. Specific mutants lacking the NH2 terminus were targeted to the cis Golgi but were not detectable in surface caveolae. Moreover, a 32-amino acid segment of the putative COOH-terminal cytoplasmic domain of caveolin-3 was targeted specifically and exclusively to the Golgi complex and could target a soluble heterologous protein, green fluorescent protein, to this compartment. Palmitoylation-deficient COOH-terminal mutants showed negligible association with the Golgi complex. This study defines unique Golgi targeting information in the caveolin molecule and identifies the cis Golgi complex as an intermediate compartment on the caveolin cycling pathway.
小窝蛋白是整合膜蛋白,是小窝的主要成分。此外,有人提出小窝蛋白在细胞内区室和细胞表面之间循环,但小窝蛋白分子的确切运输途径和靶向信息尚未明确。我们发现,针对小窝蛋白支架结构域或小窝蛋白-1羧基末端的抗体对小窝蛋白的高尔基体池具有显著特异性,通过免疫荧光无法识别表面小窝蛋白。为了更详细地分析小窝蛋白的高尔基体靶向,在成纤维细胞中表达了小窝蛋白突变体。缺乏氨基末端的特定突变体靶向顺式高尔基体,但在表面小窝中无法检测到。此外,小窝蛋白-3假定的羧基末端胞质结构域的一个32个氨基酸的片段特异性且唯一地靶向高尔基体复合体,并且可以将可溶性异源蛋白绿色荧光蛋白靶向到该区室。棕榈酰化缺陷的羧基末端突变体与高尔基体复合体的结合可忽略不计。这项研究确定了小窝蛋白分子中独特的高尔基体靶向信息,并将顺式高尔基体复合体确定为小窝蛋白循环途径上的一个中间区室。