Effects of methylcyclodextrin on lysosomes.
The cholesterol complexing agent methyl-cyclodextrin (MCD) provides an efficient mean for the removal of cholesterol from biological membranes. In order to study the effects of this agent on the lysosomal membrane in situ, we treated HepG2 cells with MCD and studied the effects of this treatment on lysosomes in isolated fractions. We found that lysosomes prepared from treated cells are more sensitive to various membrane perturbing treatments such as: incubation of lysosomes in isotonic glucose, in hypotonic sucrose or in the presence of the lytic agent glycyl-L-phenylalanine 2-naphthylamide. The lysosomal membrane is also less resistant to increased hydrostatic pressure. Centrifugation methods were used to analyse the effect of MCD on lysosomes. Isopycnic centrifugation in sucrose density gradients demonstrates that the drug induces a reversible density increase of the lysosomes. Our study indicates that extracellularly added MCD can modify the properties of the lysosomal membrane in living cells. It suggests that MCD could be an effective tool to modulate the physical properties of lysosomes within intact cells and to monitor the cellular responses to such modifications.
Jadot
M
M
Laboratory of Physiological Chemistry, University of Namur (FUNDP), Namur, Belgium. michel.jadot@fundp.ac.be
Andrianaivo
F
F
Dubois
F
F
Wattiaux
R
R
D020789:Cathepsin C / Q000378:metabolism; D002500:Centrifugation, Isopycnic; D003505:Cyclodextrins / Q000494:pharmacology*; D004151:Dipeptides / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006801:Humans; D006874:Hydrostatic Pressure; D007038:Hypotonic Solutions; D007552:Isotonic Solutions; D008247:Lysosomes / Q000187:drug effects* / Q000201:enzymology; D009997:Osmotic Pressure; D010539:Permeability; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D014407:Tumor Cells, Cultured; D047392:beta-Cyclodextrins; D001616:beta-Galactosidase / Q000378:metabolism
D003505:Cyclodextrins; D004151:Dipeptides; D007038:Hypotonic Solutions; D007552:Isotonic Solutions; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; C041865:glycylphenylalanine 2-naphthylamide; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; D001616:beta-Galactosidase; D020789:Cathepsin C; D005947:Glucose