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Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions
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Cell surface annexin II is a high affinity receptor for the alternatively spliced segment of tenascin-C

C Y Chung1, H P Erickson

  • 1Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710.

The Journal of Cell Biology
|July 1, 1994
PubMed

Insights

Tenascin-C (TN-C) binds to glioma and endothelial cells via its alternatively spliced segment (TNfnA-D). This binding is mediated by annexin II, a 35-kD protein, identified as a novel extracellular receptor for TN-C.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Soluble tenascin-C (TN-C) is an extracellular matrix glycoprotein involved in cell adhesion and migration.
  • The binding interactions of TN-C with cell surface receptors are not fully understood.
  • Alternative splicing generates different TN-C isoforms with potentially distinct functions.

Purpose of the Study:

  • To investigate the binding of soluble tenascin-C (TN-C) to various cell lines.
  • To identify the specific domains of TN-C responsible for cell binding.
  • To characterize the cell surface receptor for TN-C.

Main Methods:

  • Radioligand binding assays were used to study TN-C binding to U-251MG glioma cells, bovine aortic endothelial cells, and hamster fibroblasts.
  • Recombinant TN-C domains were employed to map binding sites.
  • Scatchard analysis quantified binding parameters.
  • Blot binding assays and affinity chromatography identified the receptor.
  • Protein sequencing and antibody blocking experiments confirmed receptor identity.

Main Results:

  • Specific binding of TN-C was observed on human glioma and bovine aortic endothelial cells, but not on hamster fibroblasts.
  • The alternatively spliced TN-C segment (TNfnA-D) strongly inhibited TN-C binding and bound directly to cells.
  • Scatchard analysis revealed approximately 2-5 x 10^5 binding sites/cell with a 2 nM dissociation constant for TNfnA-D.
  • A 35-kD cell surface protein, identified as annexin II, was found to be the receptor for TNfnA-D.
  • Annexin II directly bound TN-C and its alternatively spliced segment, and antibodies to annexin II blocked TNfnA-D binding to cells.

Conclusions:

  • Annexin II functions as an extracellular receptor for the alternatively spliced segment of tenascin-C.
  • This interaction may mediate cellular responses to soluble TN-C in the extracellular matrix.
  • The identification of annexin II as a TN-C receptor provides new insights into cell-matrix communication.

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