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Related Experiment Videos

Cell surface molecules that bind fibronectin's matrix assembly domain.

A H Limper1, B J Quade, R M LaChance

  • 1Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.

The Journal of Biological Chemistry
|May 25, 1991
PubMed
Summary
This summary is machine-generated.

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Researchers identified cell surface molecules that bind to fibronectin's matrix assembly domain. This binding is crucial for fibronectin extracellular matrix assembly by mesenchymal cells.

Area of Science:

  • Cell Biology
  • Extracellular Matrix Research
  • Protein Interactions

Background:

  • Fibronectin (FN) assembly into extracellular matrices (ECM) by mesenchymal cells requires cell interaction with specific FN sites.
  • A 29-kDa fragment of FN, containing cell-binding and amino-terminal repeats, inhibits FN matrix assembly.
  • Cell surface molecules binding to this 29-kDa fragment have been difficult to purify using standard methods.

Purpose of the Study:

  • To identify and characterize cell surface molecules that interact with the 29-kDa FN matrix assembly domain.
  • To elucidate the molecular mechanisms underlying fibronectin matrix assembly.

Main Methods:

  • Radiolabeling (125I) of the 29-kDa FN fragment.
  • Chemical cross-linking of bound 29-kDa fragment to fibroblast cell surfaces using bis(sulfosuccinimidyl) suberate.

Related Experiment Videos

  • Extraction of cross-linked complexes and analysis by SDS-PAGE.
  • Inhibition studies with unlabeled fragments and EDTA.
  • Immunoprecipitation using anti-fibronectin and anti-integrin antibodies.

    • Main Results:

    • Specific cross-linking of 125I-labeled 29-kDa fragment to cell surface molecules resulted in radiolabeled complexes of 56, 150, and 280 kDa.
    • Complex formation was inhibited by excess unlabeled 29-kDa fragment, confirming binding specificity.
    • The 280-kDa band contained 29-kDa cross-linked to cell-surface fibronectin, identified via immunoprecipitation.
    • The 150-kDa complex formation was EDTA-sensitive, suggesting a requirement for divalent cations, but was not recognized by anti-α5β1 integrin antibodies.

    Conclusions:

    • The 29-kDa FN matrix assembly domain binds to multiple cell surface molecules, including cell-surface fibronectin and a novel cation-dependent 150-kDa complex.
    • These interactions are critical for the assembly of fibronectin into the extracellular matrix.
    • Further investigation is needed to identify the precise molecular nature of the 150-kDa complex and its role in cell adhesion and matrix assembly.