Related Experiment Videos
Ion and water retention by permeabilized cells
1Department of Physiology and Molecular Biophysics, Baylor College of Medicine, Houston, TX 77030.
Scanning Microscopy
|March 1, 1988
Summary
Nonionic detergents like Triton X-100 and Brij 58 rapidly remove cell membranes. This study reveals that cellular ions and proteins are not fully dissolved in water and membrane integrity isn't essential for their retention.
Area of Science:
- Cell Biology
- Biochemistry
- Membrane Biophysics
Background:
- Nonionic detergents are used to permeabilize cell membranes.
- Understanding the behavior of intracellular components after membrane removal is crucial.
Purpose of the Study:
- To investigate the effects of nonionic detergents (Triton X-100, Brij 58) on cell membranes.
- To study the hydration, ionic asymmetry, and release kinetics of ions and proteins from membraneless cellular preparations.
Main Methods:
- Treatment of thymus lymphocytes and H-50 monolayer cells with Triton X-100 and Brij 58.
- Isolation of nuclei and study of their hydration in response to varying monovalent ionic concentrations.
- Kinetic analysis of potassium and protein release.
Main Results:
- Triton X-100 and Brij 58 rapidly removed lipid membranes (2-5 minutes).
- Nuclei hydration was sensitive to buffer ionic concentration; high concentrations caused swelling.
- Potassium and detergent-mobilized proteins showed co-compartmentalization and slow release (≥10 minutes) after Brij 58 treatment.
Conclusions:
- Most cellular potassium and diffusible proteins are not fully dissolved in the aqueous phase.
- Cell membrane integrity is not essential for retaining ions and proteins.
- Detergent-mediated cell permeabilization offers insights into intracellular solute behavior.