A novel adaptor-related protein complex.
A novel adaptor-related protein complex.
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DOI：
10.1083/jcb.133.4.749
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发表时间：
1996-05
期刊：
影响因子：
--
通讯作者：
Robinson MS
中科院分区：
文献类型：
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作者：
Simpson F;Bright NA;West MA;Newman LS;Darnell RB;Robinson MS
外壳蛋白是介导膜运输途径的运输囊泡出芽所必需的，但对于许多途径，这些蛋白质尚未被鉴定。我们已经产生了针对p47的抗体，p47是网格蛋白包被的囊泡的衔接子复合物的中链的同源物（Pevsner，J.，W. Volknandt，B.R.黄先生及R.H.舍勒。1994.基因（Amst.）。146：279-283），以确定这种蛋白质是否可能是一种新型外套的成分。p47与另外三种蛋白质共免疫沉淀：160和25 kD的两种未知蛋白质，以及β-NAP，β/β ′-衔接素的同源物，表明它是衔接子样异源四聚体复合物的亚基。然而，p47并不富集在网格蛋白包被的囊泡制剂中。招募的p47含有复合物到细胞膜上的刺激GTP γ S和布雷菲德菌素A，表明，像其他外壳蛋白，其膜协会是由ARF调节。新招募的复合物定位于与TGN相关的非网格蛋白包被的芽和囊泡。神经元细胞原代培养物中的内源性复合物也定位于TGN，此外，一些复合物与质膜相关。这些结果表明，该复合物是一种新型的外套，促进出芽的囊泡从TGN，可能是运输新合成的蛋白质到质膜的组件。
Coat proteins are required for the budding of the transport vesicles that mediate membrane traffic pathways, but for many pathways such proteins pathways, but for many pathways such proteins have not yet been identified. We have raised antibodies against p47, a homologue of the medium chains of the adaptor complexes of clathrin-coated vesicles (Pevsner, J., W. Volknandt, B.R. Wong, and R.H. Scheller. 1994. Gene (Amst.). 146:279-283), to determine whether this protein might be a component of a new type of coat. p47 coimmunoprecipitates with three other proteins: two unknown proteins of 160 and 25 kD, and beta-NAP, a homologue of the beta/beta'-adaptins, indicating that it is a subunit of an adaptor-like heterotetrameric complex. However, p47 is not enriched in preparations of clathrin-coated vesicles. Recruitment of the p47-containing complex onto cell membranes is stimulated by GTP gamma S and blocked by brefeldin A, indicating that, like other coat proteins, its membrane association is regulated by an ARF. The newly recruited complex is localized to non-clathrin-coated buds and vesicles associated with the TGN. Endogenous complex in primary cultures of neuronal cells is also localized to the TGN, and in addition, some complex is associated with the plasma membrane. These results indicate that the complex is a component of a novel type of coat that facilitates the budding of vesicles from the TGN, possibly for transporting newly synthesized proteins to the plasma membrane.