The isolation of lysosomes from normal rat liver by affinity chromatography.
Lysosomes from normal rat liver were isolated by affinity chromatography using Sepharose-bound Ricinus communis agglutinins I + II. Characterization of the lysosomal fraction by marker enzymes showed--compared with the homogenate--an enrichment in: acid phosphatase and arylsulfatase about 30- to 60-fold, the tartrate-sensitive acid phosphatase about 95-fold, whereas beta-D-glucosidase, beta-D-galactosidase and sphingomyelinase showed a much higher enrichment of 170- to 260-fold. Marker enzymes for other cell organelles were not detectable. The phospholipid pattern and optical control with electron microscopy gave further indications that the isolated fractions were very rich in lysosomes. A comparison of the phospholipid compositions of plasma membranes isolated from normal rat liver and membranes from the isolated fractions of lysosomes, showed that they were quite different; in particular bis(monoacylglycero)phosphate, which we found to be a typical lysosomal phospholipid, was absent in plasma membranes.
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000648:ultrastructure; D002846:Chromatography, Affinity / Q000379:methods; D005260:Female; D066298:In Vitro Techniques; D050356:Lipid Metabolism; D008099:Liver / Q000648:ultrastructure*; D008247:Lysosomes / Q000201:enzymology / Q000648:ultrastructure*; D008862:Microsomes, Liver / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013108:Sphingomyelin Phosphodiesterase / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000032:analysis
D011506:Proteins; D013385:Succinate Dehydrogenase; D000135:Acid Phosphatase; D013108:Sphingomyelin Phosphodiesterase