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Related Experiment Videos

Purification and partial characterization of ring-shaped miniparticles.

F R Harmon, W H Spohn, N Domae

    Cell Biology International Reports
    |May 1, 1983
    PubMed
    Summary

    Researchers purified novel ring-shaped miniparticles from human cells. These multisubunit protein particles are found in various cellular locations and exhibit immunological similarity across cell lines.

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    Area of Science:

    • Cell biology
    • Molecular biology
    • Biochemistry

    Background:

    • Small ring-shaped miniparticles are observed in the nucleus, nucleolus, and cytoplasm of diverse normal and malignant cells.
    • The precise composition and structure of these miniparticles remain incompletely characterized.

    Purpose of the Study:

    • To purify and characterize the composition and properties of the ring-shaped miniparticles.
    • To investigate the cellular localization and potential functional significance of these particles.

    Main Methods:

    • Purification involved differential and gradient centrifugation, hydroxylapatite chromatography, and preparative electrophoresis.
    • Characterization utilized one-dimensional SDS-PAGE, two-dimensional IEF/SDS gels, and sucrose gradient sedimentation (S20,W).
    • Immunological similarity was assessed across several human cell lines.

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    Main Results:

    • The miniparticles are multisubunit proteins composed of at least six polypeptide chains (Mr 20,000-30,000 Daltons).
    • These particles can be reversibly dissociated in EDTA-containing buffers, indicating a dependence on divalent cations.
    • The particles are not membrane-associated and contain no detectable carbohydrate.
    • Sedimentation analysis revealed an S20,W of 21S on sucrose gradients.
    • Immunological assays demonstrated similarity among the particles isolated from various human cell lines.

    Conclusions:

    • The study successfully purified and characterized novel ring-shaped miniparticles from human cells.
    • These particles represent a distinct class of multisubunit protein structures with a conserved immunological profile.
    • Further research is warranted to elucidate the specific functions of these miniparticles in cellular processes.