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Bovine intracellular cysteine proteinases
Acta Biologica Et Medica Germanica
|January 1, 1981
Summary
Researchers isolated bovine cathepsins B, H, and S, finding that acid pH incubation altered their enzyme activities. These cysteine proteinases showed specific inhibition patterns and substrate preferences, providing insights into their function.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Cathepsins are cysteine proteinases involved in various cellular processes.
- Understanding the properties of cathepsins B, H, and S is crucial for their study.
Purpose of the Study:
- To isolate and characterize bovine cathepsins B, H, and S.
- To investigate the effects of incubation conditions on enzyme activity.
- To determine enzyme specificity and inhibition profiles.
Main Methods:
- Isolation of cathepsins from bovine lymph nodes and spleen.
- Enzyme activity assays (BANA hydrolase, LeuNA).
- Electrophoresis and isoelectric focusing for homogeneity and isoforms.
- Circular Dichroism (CD) spectroscopy for structural analysis.
- Inhibition studies with specific reagents and venom inhibitors.
Main Results:
- Cathepsins B, H, and S were isolated and confirmed as electrophoretically homogeneous.
- Incubation at acid pH and 37°C altered BANA hydrolase and LeuNA activities, decreasing cathepsin S activity.
- Isoelectric focusing revealed multiple forms for each enzyme.
- CD spectra indicated a high percentage of unordered structure.
- Enzymes were inhibited by thiol blocking reagents, leupeptin, and Vipera ammodytes venom inhibitor.
Conclusions:
- Bovine cathepsins B, H, and S possess distinct properties regarding activity, structure, and inhibition.
- Acidic conditions significantly impact the activity of these enzymes.
- The characterized properties provide a basis for further functional studies of these cysteine proteinases.