Velasco A, Hendricks L, Moremen K W, Tulsiani D R, Touster O, Farquhar M G
Division of Cellular and Molecular Medicine and Center for Molecular Genetics, University of California, San Diego, La Jolla 92093-0651.
J Cell Biol. 1993 Jul;122(1):39-51. doi: 10.1083/jcb.122.1.39.
alpha-mannosidases I and II (Man I and II) are resident enzymes of the Golgi complex involved in oligosaccharide processing during N-linked glycoprotein biosynthesis that are widely considered to be markers of the cis- and medial-Golgi compartments, respectively. We have investigated the distribution of these enzymes in several cell types by immunofluorescence and immunoelectron microscopy. Man II was most commonly found in medial- and/or trans- cisternae but showed cell type-dependent variations in intra-Golgi distribution. It was variously localized to either medial (NRK and CHO cells), both medial and trans (pancreatic acinar cells, enterocytes), or trans- (goblet cells) cisternae, or distributed across the entire Golgi stack (hepatocytes and some enterocytes). The distribution of Man I largely coincided with that of Man II in that it was detected primarily in medial- and trans-cisternae. It also showed cell type dependent variations in its intra-Golgi distribution. Man I and Man II were also detected within secretory granules and at the cell surface of some cell types (enterocytes, pancreatic acinar cells, goblet cells). In the case of Man II, cell surface staining was shown not to be due to antibody cross-reactivity with oligosaccharide epitopes. These results indicate that the distribution of Man I and Man II within the Golgi stack of a given cell type overlaps considerably, and their distribution from one cell type to another is more variable and less compartmentalized than previously assumed.
α-甘露糖苷酶I和II(Man I和Man II)是高尔基体复合体中的驻留酶,参与N-连接糖蛋白生物合成过程中的寡糖加工,通常分别被认为是顺式和中间高尔基体区室的标志物。我们通过免疫荧光和免疫电子显微镜研究了这些酶在几种细胞类型中的分布。Man II最常见于中间和/或反式潴泡,但在高尔基体内部的分布表现出细胞类型依赖性变化。它定位于中间(NRK细胞和CHO细胞)、中间和反式(胰腺腺泡细胞、肠上皮细胞)或反式(杯状细胞)潴泡,或分布于整个高尔基体堆栈(肝细胞和一些肠上皮细胞)。Man I的分布与Man II基本一致,主要在中间和反式潴泡中检测到。它在高尔基体内部的分布也表现出细胞类型依赖性变化。在一些细胞类型(肠上皮细胞、胰腺腺泡细胞、杯状细胞)的分泌颗粒和细胞表面也检测到了Man I和Man II。就Man II而言,细胞表面染色并非由于抗体与寡糖表位的交叉反应。这些结果表明,给定细胞类型的高尔基体堆栈中Man I和Man II的分布有相当大的重叠,并且它们在不同细胞类型之间的分布比以前认为的更具变异性且分区性更小。