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[Determination of lysosome membrane stability].
Biokhimiia (Moscow, Russia)
|May 1, 1975
Summary
Triton X-100 detergent effectively releases lysosome enzymes. A new method using Triton X-100 concentrations assesses lysosome membrane stability, revealing issues in starvation, protein deficiency, and aging.
Area of Science:
- Biochemistry
- Cell Biology
- Enzymology
Background:
- Lysosomes are crucial cellular organelles containing various hydrolytic enzymes.
- Lysosome membrane integrity is vital for cellular function and is affected by physiological and pathological conditions.
- Non-ionic detergents are commonly used to study membrane-bound enzymes and membrane properties.
Purpose of the Study:
- To investigate the effect of different non-ionic detergents (Triton X-100, Triton X-305, BRIJ-35, Triton WR-1339) on rat liver lysosome enzyme activity.
- To determine the optimal Triton X-100 concentration for complete lysosome enzyme release.
- To develop a method for assessing lysosome membrane stability using differential enzyme release.
Main Methods:
- Studied the total and non-sedimentable activity of eight rat liver lysosome enzymes.
- Tested various concentrations of non-ionic detergents, focusing on Triton X-100.
- Developed a method involving sequential treatment of membrane particles with increasing Triton X-100 concentrations (0.025%, 0.05%, 0.1%) and measuring non-sedimentable enzyme activity.
Main Results:
- Triton X-100 at 0.1% and higher concentrations completely released lysosome enzymes.
- Lower Triton X-100 concentrations (0.025-0.05%) differentially released enzymes, indicating varying binding strengths.
- Acid DNase, beta-galactosidase, beta-glucuronidase, and acid phosphatase showed higher release levels at low concentrations compared to other enzymes.
- The developed method detected compromised lysosome membrane stability in rats subjected to starvation, protein deficiency, and aging.
Conclusions:
- Triton X-100 is an effective detergent for lysosome enzyme solubilization and membrane studies.
- A novel method based on differential enzyme release by increasing Triton X-100 concentrations allows for the assessment of lysosome membrane stability.
- This method is valuable for studying lysosome membrane integrity under various physiological and pathological states, including aging and nutritional deficiencies.