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Related Experiment Videos

Antibody mapping of tissue factor implicates two different exon-encoded regions in function.

W Ruf1, A Rehemtulla, T S Edgington

  • 1Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA 92037.

The Biochemical Journal
|September 15, 1991
PubMed
Summary

Tissue Factor (TF) binds Factor VII/VIIa via specific regions. Antibodies targeting these regions, located in exon 3 and exon 4, inhibit binding, revealing TF's functional requirements.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Structure-Function Relationships

Background:

  • Tissue Factor (TF) is a transmembrane glycoprotein acting as the cellular receptor for Factor VII and Factor VIIa (VIIa).
  • TF facilitates the catalytic activity of VIIa within the TF:VIIa complex, crucial for hemostasis.

Purpose of the Study:

  • To identify the specific structural regions (epitopes) on Tissue Factor responsible for binding Factor VII and VIIa.
  • To investigate the functional significance of these identified TF regions in ligand recognition and binding affinity.

Main Methods:

  • Utilized monoclonal antibodies (MAbs) and sequence-specific polyclonal antibodies against native TF.
  • Analyzed antibody inhibition of Factor VII binding to TF.
  • Localized antibody epitopes using reciprocal competition, proteolytic TF fragments, and C-terminal deletional mutations.

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Main Results:

  • Identified two distinct epitopes on TF involved in Factor VII/VIIa binding.
  • Localized one epitope (locus II) to residues Thr40-Val83 (exon 3) and another (locus I) to fragments including exon 4 (Thr106-Lys165).
  • Antibodies targeting linear sequences within these exons also inhibited Factor VII binding.

Conclusions:

  • Two specific regions of Tissue Factor, encoded by exon 3 and exon 4, are essential for high-affinity binding of Factor VII/VIIa.
  • These findings elucidate the structural basis of TF's cofactor function in ligand recognition.