Sensitivity of lysosomal enzymes to the plant alkaloid sanguinarine: comparison with other SH-specific agents.
The influence of the benzo[c]phenanthridine alkaloid sanguinarine on some lysosomal enzyme activities was investigated. Sanguinarine inhibits lysosomal hydrolases in homogenates of cultured mouse fibroblasts. After incubation of mouse fibroblasts in culture with 100 microM sanguinarine an approximately 50% decrease in the activities of N-acetyl-beta,D-glucosaminidase (NAGA), beta-galactosidase (GAL), arylsulfatase and acid lipase was observed. Because the biological activity of sanguinarine might arise from the interaction of its iminium cation with enzyme thiol groups, we compared its effect on NAGA, GAL and acid phosphatase (AcP) activities with the effects of SH-specific reagents p-chloromercuribenzoic acid (CPMA) and N-ethylmaleimide (NEM). Treatment of lysosomal fractions with millimolar concentrations of sanguinarine induces a dose-dependent inhibition of the enzymes; for example, 0.6 mM sanguinarine causes approximately a 40% decrease in AcP and NAGA activities. NEM has similar effects, and increasing the preincubation temperature from 0 degrees C to 37 degrees C intensifies the inhibition due to both agents. CPMA also inhibits the activity of GAL (IC50 0.7 microM), AcP (IC50 12.5 microM) and NAGA (IC50 6.8 microM) in a dose-dependent manner but is more potent than sanguinarine or NEM. Comparative analysis of the primary structures of these enzymes using the program BLAST reveals the presence of highly conserved cysteine residues, which confirms the importance of thiol-groups for their activities. Thus, both the experimental observations obtained in this study and the literature data imply a significant role of redox-based mechanisms in regulating lysosomal functional activity.
Institute of Cytology, Russian Academy of Sciences, St Petersburg, Russia.
T
T
Belyaeva
E
E
Leontieva
A
A
Shpakov
T
T
Mozhenok
M
M
Faddejeva
D000118:Acetylglucosaminidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000135:Acid Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D001192:Arylsulfatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D053119:Benzophenanthridines; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006867:Hydrolases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D007546:Isoquinolines; D008049:Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D001616:beta-Galactosidase / Q000187:drug effects / Q000378:metabolism; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
D000470:Alkaloids; D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; D007546:Isoquinolines; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D020245:p-Chloromercuribenzoic Acid; C005705:sanguinarine; D006867:Hydrolases; D008049:Lipase; D000135:Acid Phosphatase; D001192:Arylsulfatases; D001616:beta-Galactosidase; D000118:Acetylglucosaminidase; D003545:Cysteine; D005033:Ethylmaleimide