Allan V J, Kreis T E
J Cell Biol. 1986 Dec;103(6 Pt 1):2229-39. doi: 10.1083/jcb.103.6.2229.
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 approximately 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. A 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.
一种针对微管相关蛋白2（MAP - 2）产生的单克隆抗体（M3A5），在多种非神经组织培养细胞中识别出一种与高尔基体复合物相关的抗原。在双重免疫荧光研究中，即使在用莫能菌素或诺考达唑破坏高尔基体结构后，M3A5染色仍与特定高尔基体标志物的染色非常相似。M3A5在培养细胞提取物的免疫印迹中识别出一条分子量约为110,000的条带；这种蛋白质被命名为110K，在从大鼠肝脏制备的高尔基体堆叠组分中含量丰富。已证明110K蛋白可通过用Triton X - 114提取富含高尔基体的组分而分配到水相中，并在pH 11.0的碳酸盐洗涤后洗脱。因此，它可能是一种外周膜蛋白。用蛋白酶K处理分离的高尔基体堆叠组分，无论有无Triton X - 100，都会导致110K蛋白完全被消化。由于110K蛋白在体外完整囊泡中可被蛋白酶作用，推测它在体内位于高尔基体膜的细胞质面上。110K蛋白在体外能够与紫杉醇聚合的微管特异性相互作用。这些结果表明，110K蛋白可能起到将高尔基体与微管网络连接的作用，因此可能属于一类新型蛋白质：微管结合蛋白。
