Pfeffer S R, Kelly R B
Cell. 1985 Apr;40(4):949-57. doi: 10.1016/0092-8674(85)90355-1.
Coated vesicles have been purified in the past on the basis of their remarkably homogeneous structure, not their function. We have succeeded in isolating two subpopulations of bovine brain coated vesicles that carry specific "cargoes," in this case two synaptic vesicle membrane polypeptides (Mr = 95,000 and 65,000). Monoclonal antibodies that recognize cytoplasmic domains of these polypeptides can penetrate the clathrin coat and recognize them on the outer surface of the coated vesicle membrane. An immunoadsorption technique could therefore be used to fractionate coated vesicles on the basis of their membrane composition. The subpopulations have the normal complement of conventional coated vesicle proteins. Exclusive, however, to the subpopulations that carry synaptic vesicle polypeptides are two new coated vesicle polypeptides (Mr = 38,000 and 29,000).
过去,有被小泡是根据其极为均一的结构而非功能进行纯化的。我们成功分离出了牛脑有被小泡的两个亚群,它们携带特定的“货物”,在这种情况下是两种突触小泡膜多肽(分子量分别为95,000和65,000)。识别这些多肽胞质结构域的单克隆抗体能够穿透网格蛋白衣被,并在有被小泡膜的外表面识别它们。因此,可以使用免疫吸附技术根据有被小泡的膜组成对其进行分级分离。这些亚群具有常规有被小泡蛋白的正常组成。然而,携带突触小泡多肽的亚群特有的是两种新的有被小泡多肽(分子量分别为38,000和29,000)。