The Isolation of Lysosomes from Normal Rat Liver by Affinity Chromatography

Publisher Website
Google Scholar
Abstract
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 EM 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 different; in particular bis(monoacylglycero)phosphate, which was a typical lysosomal phospholipid, was absent in plasma membranes.