Parenti G, Willemsen R, Hoogeveen A T, Verleun-Mooyman M, Van Dongen J M, Galjaard H
Eur J Cell Biol. 1987 Feb;43(1):121-7.
This study represents the first example of immunological localization of lysosomal acid phosphatase. The intracellular localization of lysosomal acid phosphatase was investigated with immunocytochemical methods at the light and electron microscopical level in cultured fibroblasts obtained from normal subjects and from a patient with I-cell disease. Double-labeling studies using fluorescence microscopy showed that acid phosphatase is present in the same organelles as other hydrolases. At the electron microscopic level in control fibroblasts acid phosphatase was found in the rough endoplasmic reticulum, lysosomes, at the plasma membrane, in vesicles just below the plasma membrane and in multivesicular bodies. This localization was comparable with that of other lysosomal enzymes tested (acid alpha-glucosidase, N-acetyl-beta-hexosaminidase, beta-galactosidase). Acid phosphatase labeling was mainly found in association with the lysosomal membrane and with membranous material present within the lysosome. In I-cell fibroblasts the label was present in the same subcellular organelles but always associated with membranous structures. We suggest that the association of acid phosphatase with membranes might explain the normal enzyme activity found in I-cell fibroblasts.
本研究是溶酶体酸性磷酸酶免疫定位的首个实例。运用免疫细胞化学方法，在光学显微镜和电子显微镜水平上，对取自正常受试者和一名患I-细胞病患者的培养成纤维细胞中溶酶体酸性磷酸酶的细胞内定位进行了研究。利用荧光显微镜进行的双重标记研究表明，酸性磷酸酶与其他水解酶存在于相同的细胞器中。在对照成纤维细胞的电子显微镜水平下，酸性磷酸酶存在于粗面内质网、溶酶体、质膜、质膜下方的小泡以及多泡体中。这种定位与所检测的其他溶酶体酶（酸性α-葡萄糖苷酶、N-乙酰-β-己糖胺酶、β-半乳糖苷酶）的定位相当。酸性磷酸酶标记主要发现与溶酶体膜以及溶酶体内的膜状物质相关。在I-细胞成纤维细胞中，标记存在于相同的亚细胞细胞器中，但始终与膜状结构相关。我们认为酸性磷酸酶与膜的关联可能解释了在I-细胞成纤维细胞中发现的正常酶活性。
