Two sites in the tissue factor extracellular domain mediate the recognition of the ligand factor VIIa.
Tissue factor (TF) binds the serine protease coagulation factor VIIa and initiates the coagulation protease cascade by forming a catalytic cofactor-enzyme complex. Using a photoactivatable crosslinking reagent coupled to factor VIIa, we have identified interactive sites in the amino-terminal (residues 44-84) and the carboxyl-terminal (residues 129-169) aspect of the extracellular domain of TF. Epitopes of inhibitory antibodies have previously indicated participation of these regions in TF function. The presence of the gamma-carboxyglutamic acid domain in factor VIIa appears to facilitate the interaction with the negatively charged, amino-proximate site, whereas crosslinking of TF with VIIa or des-(1-38)-VIIa at the positively charged carboxyl-proximate site was similar. Lack of alpha-helical secondary structure in the TF extracellular domain is consistent with the proposed structural similarity of TF with the cytokine receptor family. The interactive sites identified for TF are located in sequence spans that demonstrate a low degree of sequence conservation among the members of this receptor family. Regions with highly conserved residues, such as sequences encoded by exon 2 and 5 in TF, were not implicated in ligand recognition, suggesting that conserved residues in the receptor family may maintain the common beta-strand architecture, and variable regions provide a pair of nonidentical motifs for oriented ligand recognition.
Department of Immunology, Scripps Research Institute, La Jolla, CA 92037.
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D000345:Affinity Labels; D000595:Amino Acid Sequence; D001665:Binding Sites; D002942:Circular Dichroism; D003432:Cross-Linking Reagents; D005110:Extracellular Space; D015942:Factor VIIa / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D010777:Photochemistry; D011487:Protein Conformation; D011994:Recombinant Proteins; D016415:Sequence Alignment; D013925:Thromboplastin / Q000737:chemistry / Q000378:metabolism*
D000345:Affinity Labels; D003432:Cross-Linking Reagents; D008024:Ligands; D011994:Recombinant Proteins; D013925:Thromboplastin; D015942:Factor VIIa