大鼠肝脏中溶酶体β-半乳糖苷酶的免疫细胞化学定位
Immunocytochemical localization of lysosomal beta-galactosidase in rat liver.
作者信息
Novikoff P M, La Russo N F, Novikoff A B, Stockert R J, Yam A, Le Sage G D
出版信息
J Cell Biol. 1983 Nov;97(5 Pt 1):1559-65. doi: 10.1083/jcb.97.5.1559.
Abstract
beta-galactosidase is a ubiquitous lysosomal hydrolase that specifically cleaves terminal beta-galactosyl residues from glycoproteins, glycosaminoglycans, oligosaccharides, and glycolipids. To study the intracellular distribution of this enzyme, we prepared a specific polyclonal antibody to lysosomal beta-galactosidase by immunizing rabbits with a highly purified preparation of beta-galactosidase from rat liver. Using this antibody we employed an immunocytochemical technique (protein A coupled to horseradish peroxidase and diaminobenzidine cytochemistry) and showed that beta-galactosidase is present in all hepatocytes of the rat liver. All types of lysosomes, the rough endoplasmic reticulum, and the specialized region of smooth endoplasmic reticulum known as GERL showed immunoreactivity. This in situ distribution suggests that these organelles are involved in the biosynthesis and intracellular sorting of this lysosomal enzyme.
摘要
β-半乳糖苷酶是一种普遍存在的溶酶体水解酶，它能特异性地从糖蛋白、糖胺聚糖、寡糖和糖脂上切割末端β-半乳糖基残基。为了研究这种酶在细胞内的分布，我们用大鼠肝脏高度纯化的β-半乳糖苷酶制剂免疫兔子，制备了针对溶酶体β-半乳糖苷酶的特异性多克隆抗体。使用这种抗体，我们采用免疫细胞化学技术（蛋白A偶联辣根过氧化物酶和二氨基联苯胺细胞化学），结果表明β-半乳糖苷酶存在于大鼠肝脏的所有肝细胞中。所有类型的溶酶体、粗面内质网以及被称为GERL的滑面内质网特殊区域均显示出免疫反应性。这种原位分布表明这些细胞器参与了这种溶酶体酶的生物合成和细胞内分选。
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