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Related Experiment Videos

Actin-severing activity copurifies with phosphofructokinase.

A Füchtbauer, B M Jockusch, E Leberer

    Proceedings of the National Academy of Sciences of the United States of America
    |December 1, 1986
    PubMed
    Summary
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    Muscle 6-phosphofructokinase (PFK) disintegrates cellular stress fibers, similar to the actin-severing protein brevin. This suggests novel cellular control mechanisms linking microfilament function and glycolysis.

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Background:

    • Microfilament bundles, or stress fibers, are crucial for cellular structure and function.
    • Actin-severing proteins like gelsolin and brevin play roles in regulating the cytoskeleton.
    • The enzyme 6-phosphofructokinase (PFK) is primarily known for its role in glycolysis.

    Purpose of the Study:

    • To investigate the potential role of muscle 6-phosphofructokinase (PFK) in regulating cellular microfilament organization.
    • To compare the effects of PFK on stress fibers with known actin-severing proteins.

    Main Methods:

    • Microinjection of purified muscle PFK into tissue culture cells.
    • Experiments using detergent-extracted and ethanol-fixed cells.
    • Calcium-dependency assays for stress fiber disruption.

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  • Biochemical analysis including SDS-PAGE and Western blotting using antibodies.

    • Main Results:

    • Microinjection of PFK caused reversible disintegration of stress fibers.
    • PFK's disruption of stress fibers mimicked the effects of brevin, an actin-severing protein.
    • PFK disrupted stress fibers in a calcium-dependent manner in both live and fixed cells.
    • PFK shared biochemical and immunological properties with gelsolin, including comigration and antibody cross-reactivity.

    Conclusions:

    • Muscle PFK exhibits actin-severing capabilities, influencing microfilament organization.
    • PFK's interaction with the cytoskeleton suggests a direct link between glycolysis and cellular structure.
    • These findings reveal unexpected cellular control mechanisms involving PFK in both microfilament dynamics and metabolic pathways.