The isolation of lysosomes from Ehrlich ascites tumor cells following pretreatment of mice with Triton WR-1339.
A method is described for obtaining highly purified lysosomes from Ehrlich ascites tumo cells grown in mice injected with Triton WR-1339. The isolated particles show a high specific activity for aryl sulfatase, representing an 80-90-fold purification over the homogenate, and a 15-18% yield of the total enzyme activity. Mitochondrial and microsomal marker enzymes are present in negligible amounts (0.2% of the activity of the homogenate). The biochemical evidence for a rather high degree of homogeneity of the fraction is supported by the electron microscopic examination of the purified lysosomes. The intracellular localizations of N-acetyl-beta-glucosaminidase, NADH-cytochrome c reductase and NADPH-cytochrome c reductase in Ehrlich ascites cells are also reported, the first two being present in highest concentration in the combined mitochondrial-lysosomal fraction and the third in the microsomal fraction.
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology / Q000473:pathology*; D002499:Centrifugation, Density Gradient; D006026:Glycoside Hydrolases / Q000032:analysis*; D006651:Histocytochemistry; D007274:Injections, Intraperitoneal; D008247:Lysosomes / Q000201:enzymology*; D008722:Methods; D008854:Microscopy, Electron; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D010088:Oxidoreductases / Q000032:analysis*; D013501:Surface-Active Agents
D013501:Surface-Active Agents; D010088:Oxidoreductases; D006026:Glycoside Hydrolases