Anderson R G, Pathak R K
Cell. 1985 Mar;40(3):635-43. doi: 10.1016/0092-8674(85)90212-0.
Recently we demonstrated that low-pH compartments can be visualized with the electron microscope using a basic congener of dinitrophenol, 3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine (DAMP), which concentrates in acidic compartments and can be detected by immunocytochemistry with a monoclonal anti-dinitrophenol antibody. We now report that DAMP also accumulates in cisternae and vesicles associated with the trans face of the Golgi apparatus. DAMP rapidly leaves this compartment when cells are incubated with the ionophore monensin, which indicates that accumulation is due to the acidic pH in this compartment. Using indirect protein A-gold immunocytochemistry, we localized fibronectin, a major secretory protein in fibroblasts, to the trans Golgi vesicles that took up DAMP. Therefore, the trans cisternae of the Golgi apparatus and forming secretory vesicles have an acidic pH.
最近我们证明,使用二硝基苯酚的一种碱性同系物3-(2,4-二硝基苯胺基)-3'-氨基-N-甲基二丙胺(DAMP),可以在电子显微镜下观察到低pH区室,DAMP会聚集在酸性区室中,并且可以通过用单克隆抗二硝基苯酚抗体进行免疫细胞化学检测到。我们现在报告,DAMP也会积聚在与高尔基体反面相关的潴泡和小泡中。当细胞与离子载体莫能菌素一起孵育时,DAMP会迅速离开这个区室,这表明积聚是由于该区室的酸性pH所致。使用间接蛋白A-金免疫细胞化学,我们将成纤维细胞中的一种主要分泌蛋白纤连蛋白定位到摄取DAMP的反式高尔基体小泡中。因此,高尔基体的反式潴泡和正在形成的分泌小泡具有酸性pH。