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Bacterial cell envelopes with functional flagella

M Eisenbach, J Adler

    The Journal of Biological Chemistry
    |August 25, 1981
    PubMed
    Summary
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    Researchers isolated bacterial cell envelopes to study flagellar rotation. Adding electron donors restored rotation, suggesting proton motive force drives it, but clockwise rotation may need cytoplasmic factors.

    Area of Science:

    • Microbiology
    • Cell Biology
    • Biochemistry

    Background:

    • Bacterial chemotaxis is controlled by flagellar rotation direction.
    • Understanding this mechanism requires a controllable system.
    • Isolating functional subcellular components is challenging.

    Purpose of the Study:

    • To isolate a flagellated, subcellular bacterial system for studying flagellar rotation control.
    • To determine the energy requirements for restoring flagellar rotation in isolated cell envelopes.
    • To investigate the role of cytoplasmic components in flagellar rotation direction.

    Main Methods:

    • Isolation of bacterial cell envelopes from Escherichia coli and Salmonella typhimurium via osmotic lysis.
    • High-intensity light microscopy to visualize individual flagella.

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  • Tethering of cell envelopes to observe flagellar motion.
  • Addition of artificial electron donors to assess energy requirements.

    • Main Results:

    • Isolated cell envelopes retained flagella and structural integrity.
    • Addition of artificial electron donors restored flagellar rotation.
    • Proton electrochemical potential appears to be the primary energy source for rotation.
    • Restored rotation was predominantly counterclockwise, unlike in intact bacteria.

    Conclusions:

    • Bacterial cell envelopes can be isolated as a functional system for studying flagellar motor function.
    • Flagellar rotation is driven by proton electrochemical potential, independent of cytoplasmic components.
    • A cytoplasmic factor may be necessary for enabling clockwise flagellar rotation, crucial for chemotaxis.