Heterogeneity of lysosomes in human fibroblasts.
Lysosomes are defined traditionally with the marker enzyme acid phosphatase. We showed recently that lysosomes from human fibroblasts can be separated into a light and dense fraction as well as prelysosomal population. We now provide evidence that although acid phosphatase is enriched in all three fractions, the marker enzyme in the prelysosomal compartment is qualitatively distinct from that of the lysosomes. Ultrastructural analysis showed that the acid phosphatase in the prelysosomal vesicles deposited an extremely electron-dense reaction product, entirely obliterating the lumen of the vesicle, in contrast to that of the light and dense lysosomes which deposited a fine and diffuse product scattered throughout the luminal space. Biochemical analysis showed that only 51% of the acid phosphatase in the prelysosomes was inhibited by tartrate, while 80% of that in the lysosomes was tartrate-inhibitable. Immunoprecipitation with antibodies specific for various isozymes of acid phosphatase showed that 39% of the acid phosphatase in the prelysosomes was of the 'lysosomal' type whereas over 50% of the acid phosphatase in the lysosomes was of this type. These results showed that acid phosphatase in the prelysosomes of human cultured fibroblasts can be distinguished from that of the lysosomes cytochemically, biochemically, and immunologically and that lysosomes, as marked by acid phosphatase, are a heterogeneous organelle.
Department of Pediatrics, McMaster University, Hamilton, Ontario, Canada.
B M
BM
Kelly
A
A
Waheed
R
R
Van Etten
P L
PL
Chang
D000135:Acid Phosphatase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000276:immunology; D002458:Cell Fractionation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000032:analysis / Q000201:enzymology* / Q000648:ultrastructure; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology / Q000648:ultrastructure; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008247:Lysosomes / Q000032:analysis / Q000145:classification / Q000201:enzymology* / Q000648:ultrastructure; D011233:Precipitin Tests; D013644:Tartrates / Q000494:pharmacology
D007527:Isoenzymes; D013644:Tartrates; D000135:Acid Phosphatase; C029768:tartaric acid