A MICROTUBULE-BINDING PROTEIN ASSOCIATED WITH MEMBRANES OF THE GOLGI-APPARATUS
A MICROTUBULE-BINDING PROTEIN ASSOCIATED WITH MEMBRANES OF THE GOLGI-APPARATUS
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DOI：
10.1083/jcb.103.6.2229
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发表时间：
1986-12-01
影响因子：
7.8
通讯作者：
KREIS, TE
中科院分区：
文献类型：
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作者：
ALLAN, VJ;KREIS, TE
针对微管相关蛋白2（MAP-2）的单克隆抗体（M3 A5）识别了多种非神经元组织培养细胞中与高尔基复合体相关的抗原。在双重免疫荧光研究中，M3 A5染色与特异性高尔基体标记非常相似，即使在用莫能菌素或诺考达唑破坏高尔基体组织后也是如此。M3 A5识别出一条Mr. apprx带。110，000;这种蛋白质，命名为110 K，富集在从大鼠肝脏制备的高尔基体组分中。110 K蛋白质已被证明分配到水相的Triton X-114提取高尔基体富集的馏分，并在pH 11.0碳酸盐洗涤后洗脱。因此，它可能是一种外周膜蛋白。蛋白酶K处理的一个孤立的高尔基堆，馏分导致完全消化的110 K蛋白质，在Triton X-100的存在和不存在。由于110 K蛋白在体外完整囊泡中可被蛋白酶接近，因此推测其在体内位于高尔基体膜的细胞质面上。110 K蛋白能够在体外与紫杉醇聚合的微管特异性相互作用。这些结果表明，110 K蛋白可能是连接高尔基体的微管网络，因此可能属于一类新的蛋白质;微管结合蛋白。
A monoclonal antibody (M3A5), raised against microtubule-associated protein 2 (MAP-2), recognized an antigen associated with the Golgi complex in a variety of non-neuronal tissue culture cells. In double immunofluorescence studies M3A5 staining was very similar to that of specific Golgi markers, even after disruption of the Golgi apparatus organization with monensin or nocodazole. M3A5 recognized one band of Mr .apprx. 110,000 in immunoblots of culture cell extracts; this protein, designated 110K, was enriched in Golgi stack fractions prepared from rat liver. The 110K protein has been shown to partition into the aqueous phase by Triton X-114 extraction of a Golgi-enriched fraction and was eluted after pH 11.0 carbonate washing. It is therefore likely to be a peripheral membrane protein. Proteinase K treatment of an isolated Golgi stack, fraction resulted in complete digestion of the 110K protein, both in the presence and absence of Triton X-100. As the 110K protein is accessible to protease in intact vesicles in vitro, it is presumably located on the cytoplasmic face of the Golgi membrane in vivo. The 110K protein was able to interact specifically with taxol-polymerized microtubules in vitro. These results suggest that the 110K protein may serve to link the Golgi apparatus to the microtubule network and so may belong to a novel class of proteins; the microtubule-binding proteins.