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Related Experiment Videos

Reconstruction of glutamine synthetase using computer averaging.

J Frank, W Goldfarb, D Eisenberg

    Ultramicroscopy
    |January 1, 1978
    PubMed
    Summary

    Researchers reconstructed the axial projection of glutamine synthetase using a novel correlation search and averaging method. The resulting sixfold symmetric radial pattern revealed molecular handedness, likely due to staining variations.

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    Area of Science:

    • Biochemistry
    • Structural Biology
    • Electron Microscopy

    Background:

    • Glutamine synthetase is a crucial enzyme in nitrogen metabolism.
    • Understanding its molecular structure is key to elucidating its function.
    • Previous structural studies may have limitations in resolution or methodology.

    Purpose of the Study:

    • To reconstruct the axial projection of the glutamine synthetase molecule.
    • To apply a new correlation search and averaging technique for structural analysis.
    • To investigate the symmetry and handedness of the glutamine synthetase structure.

    Main Methods:

    • Utilized electron micrographs of stained glutamine synthetase preparations.
    • Implemented a novel correlation search and averaging method.

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  • Analyzed the averaged structure of approximately 50 individual molecules.

    • Main Results:

    • Successfully reconstructed the axial projection of the glutamine synthetase molecule.
    • The averaged structure exhibited a radial pattern with sixfold symmetry.
    • Observed handedness in the averaged structure was identified.

    Conclusions:

    • The new method enables detailed structural reconstruction of enzymes like glutamine synthetase.
    • The observed sixfold symmetry is a key feature of the glutamine synthetase molecular architecture.
    • Nonuniformity in negative staining is proposed as the cause for the observed molecular handedness.