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A density label for membranes.

C F Fox, J H Law, N Tsukagoshi

    Proceedings of the National Academy of Sciences of the United States of America
    |October 1, 1970
    PubMed
    Summary
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    Bromostearic acid fulfills the essential fatty acid needs for Escherichia coli K12 auxotrophs. Brominated fatty acids incorporated into cell membranes increased their density, enabling separation from unbrominated membranes.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Microbiology

    Background:

    • Escherichia coli K12 requires unsaturated fatty acids for growth.
    • Bromostearic acid is a synthetic fatty acid analog.

    Purpose of the Study:

    • To determine if bromostearic acid can satisfy the essential fatty acid requirement of an unsaturated fatty acid auxotroph of Escherichia coli K12.
    • To characterize the effects of bromostearic acid incorporation on Escherichia coli K12 cell membrane properties.

    Main Methods:

    • Gas chromatography and mass spectrometry were used to analyze fatty acids from cell lipids.
    • Equilibrium centrifugation in sucrose gradients was employed to separate cell membranes based on density.

    Main Results:

    Related Experiment Videos

  • Bromostearic acid supplementation enabled growth of the Escherichia coli K12 unsaturated fatty acid auxotroph.
  • Over 55% of fatty acids in supplemented cells contained bromine.
  • Cytoplasmic membranes from bromostearate-grown cells showed a density increase of approximately 0.06 g/cm(3) compared to controls.
  • Complete separation of membranes containing brominated fatty acids was achieved.

    • Conclusions:

    • Bromostearic acid can serve as an essential fatty acid for Escherichia coli K12.
    • Incorporation of brominated fatty acids into cell membranes alters their biophysical properties, specifically increasing density.
    • Density differences allow for effective separation of membranes with and without brominated fatty acids.