The gamma-carboxyglutamic acid domain of human factor VIIa is essential for its interaction with cell surface tissue factor. 1990

T Sakai, and T Lund-Hansen, and L Thim, and W Kisiel
Blood Systems Research Foundation Laboratory, Department of Pathology, University of New Mexico School of Medicine, Albuquerque 87131.

Previous studies indicated that both plasma-derived and recombinant human factor VIIa specifically interacted with tissue factor on the surface of a human bladder carcinoma cell line (J82). In the presence of calcium ions, factor VIIa interacted with approximately 300,000 binding sites/cell with a dissociation constant (Kd) of 3.25 nM (Sakai, T., Lund-Hansen, T., Paborsky, L., Pedersen, A. H., and Kisiel, W. (1989) J. Biol. Chem. 264, 9980-9988). In this study, we compare recombinant human factor VIIa and a preparation of recombinant human factor VIIa lacking the gamma-carboxyglutamic acid domain (GD-rVIIa) with respect to their interaction with J82 cell surface tissue factor. Interaction of GD-rVIIa with J82 monolayers at 37 degrees C was specific, saturable, and exhibited a hyperbolic profile. Scatchard plots of the binding data obtained at 37 degrees C indicated a single class of binding sites for GD-rVIIa with a Kd value of 2.5 nM. GD-rVIIa interacted with about 10,000 binding sites/cell. In contrast to the tissue factor-specific binding observed for intact factor VIIa, specific binding of GD-rVIIa to the J82 cell surface was neither influenced by calcium nor blocked by prior incubation of the cells with polyclonal anti-tissue factor apoprotein IgG. In addition, cell-bound GD-rVIIa failed to activate human factor X. These results indicate that the gamma-carboxyglutamic acid domain of factor VIIa is essential for its interaction with cell surface tissue factor.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D001749 Urinary Bladder Neoplasms Tumors or cancer of the URINARY BLADDER. Bladder Cancer,Bladder Neoplasms,Cancer of Bladder,Bladder Tumors,Cancer of the Bladder,Malignant Tumor of Urinary Bladder,Neoplasms, Bladder,Urinary Bladder Cancer,Bladder Cancers,Bladder Neoplasm,Bladder Tumor,Cancer, Bladder,Cancer, Urinary Bladder,Neoplasm, Bladder,Neoplasm, Urinary Bladder,Tumor, Bladder,Tumors, Bladder,Urinary Bladder Neoplasm
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D005170 Factor X Storage-stable glycoprotein blood coagulation factor that can be activated to factor Xa by both the intrinsic and extrinsic pathways. A deficiency of factor X, sometimes called Stuart-Prower factor deficiency, may lead to a systemic coagulation disorder. Autoprothrombin III,Coagulation Factor X,Stuart Factor,Stuart-Prower Factor,Blood Coagulation Factor X,Factor 10,Factor Ten,Stuart Prower Factor,Factor X, Coagulation
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein

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