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Related Experiment Videos

Cell surface-associated proteins which bind native type IV collagen or gelatin.

M Kurkinen, A Taylor, J I Garrels

    The Journal of Biological Chemistry
    |May 10, 1984
    PubMed
    Summary

    Researchers identified four gelatin-binding polypeptides in mouse embryo cells, with the Mr = 47,000 glycoprotein binding type IV collagen. These cell surface proteins are not collagen fragments and may play roles in vivo.

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    Area of Science:

    • Cell Biology
    • Biochemistry
    • Extracellular Matrix Research

    Background:

    • Parietal endoderm cells synthesize type IV procollagen, a key component of basement membranes.
    • Understanding the interactions of cell surface proteins with extracellular matrix components is crucial for cell biology.

    Purpose of the Study:

    • To isolate and characterize gelatin-binding polypeptides from mouse embryo parietal endoderm cells.
    • To investigate the nature and potential cell surface association of these polypeptides.

    Main Methods:

    • Affinity chromatography using gelatin-Sepharose to isolate [35S]methionine-labeled polypeptides.
    • Pulse-chase experiments and [3H]mannose labeling to assess protein synthesis and glycosylation.
    • Tunicamycin sensitivity assays to confirm N-linked oligosaccharide chains.

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  • Two-dimensional gel electrophoresis to analyze isoelectric points.
  • Lactoperoxidase-catalyzed iodination to determine cell surface localization.

    • Main Results:

    • Four gelatin-binding polypeptides (Mr = 47,000, 56,000, 62,000, and 65,000) were isolated.
    • The Mr = 47,000 polypeptide is the major gelatin-binding component in cell lines synthesizing type IV procollagen.
    • Polypeptides contain N-linked oligosaccharide chains and are associated with the cell surface.
    • Only the Mr = 47,000 glycoprotein binds to native type IV collagen.

    Conclusions:

    • The identified polypeptides are distinct cell surface proteins, not derived from proteolytic cleavage of larger precursors.
    • The Mr = 47,000 glycoprotein's specific binding to type IV collagen suggests a role in cell-matrix interactions.
    • Further investigation into the in vivo functions of these cell surface components is warranted.