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Glycolysis in permeabilized L-929 cells
1Bodega Marine Laboratory, University of California 94923.
The Biochemical Journal
|October 1, 1988
Summary
Dextran sulphate permeabilized (DSP) cells maintain glycolysis. Despite plasma membrane lesions allowing large protein entry, significant protein loss is prevented, suggesting intracellular enzyme organization.
Area of Science:
- Cell Biology
- Biochemistry
- Metabolic Pathways
Background:
- Cellular metabolism relies on organized enzyme complexes.
- Understanding cytosol composition is key to cellular function.
Purpose of the Study:
- Investigate glycolysis in permeabilized mouse L-929 cells (DSP cells).
- Characterize plasma membrane integrity and protein exchange in DSP cells.
- Explore the implications of enzyme organization on cellular metabolic activity.
Main Methods:
- Utilized dextran sulphate to permeabilize L-929 cells.
- Assessed glycolytic rates under various conditions.
- Employed electron microscopy for ultrastructural analysis.
- Quantified protein and dextran infiltration using radiolabeled and fluorescent tracers.
Main Results:
- DSP cells exhibited linear glycolysis when supplied with glucose, ATP, and NAD+.
- Electron microscopy showed altered nuclear and mitochondrial ultrastructure.
- Plasma membrane lesions permitted passage of proteins up to 400 kDa.
- Despite membrane permeability, only ~10% of cellular protein was lost within 30 minutes.
Conclusions:
- Cellular enzyme organization, particularly for glycolytic enzymes, likely restricts protein efflux.
- The cytosol is not a simple aqueous solution but a highly organized compartment.
- Metabolic organization plays a crucial role in maintaining cellular integrity and function.