Sensitivity of lysosomal enzymes to the plant alkaloid sanguinarine: comparison with other SH-specific agents
| Autor: | E. A. Leontieva, Mozhenok Tp, A. O. Shpakov, T. N. Belyaeva, M. Faddejeva |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Hydrolases
Acid Phosphatase p-Chloromercuribenzoic Acid Dithiothreitol Cell Line chemistry.chemical_compound Mice Alkaloids Acetylglucosaminidase Animals Sanguinarine Cysteine Sulfhydryl Compounds Enzyme Inhibitors Arylsulfatases chemistry.chemical_classification Benzophenanthridines biology Dose-Response Relationship Drug Chemistry Alkaloid N-Ethylmaleimide Sulfhydryl Reagents Acid phosphatase Biological activity Cell Biology General Medicine Lipase Isoquinolines beta-Galactosidase Enzyme Biochemistry Ethylmaleimide biology.protein Lysosomes Oxidation-Reduction |
| Zdroj: | Cell biology international. 27(11) |
| ISSN: | 1065-6995 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text