Phospholipid-independent and -dependent interactions required for tissue factor receptor and cofactor function.
Membrane anchoring of tissue factor (TF), the cell receptor for coagulation factor VIIa (VIIa), exemplifies an effective mechanism to localize proteolysis at the cell surface. A recombinant TF mutant (TF1-219), deleted of membrane spanning and intracellular domains, was used to evaluate the role of phospholipid interactions for assembly of substrate with the catalytic TF.VIIa complex. TF1-219 was secreted by cells rather than expressed as a cell membrane protein. Unlike free VIIa, TF1-219 as well as the TF1-219.VIIa complex demonstrated no stable association with phospholipid. In the absence of lipid, kinetic evaluation of substrate factor X cleavage by free VIIa, TF.VIIa, and TF1-219.VIIa suggests that the catalytic function of VIIa rather than substrate recognition is enhanced by complex formation. Furthermore, compared with free factor X, factor X on phospholipid was preferentially cleaved as a substrate by TF1-219.VIIa. TF-dependent initiation of the coagulation protease cascades thus involves an enhancement of the activation of factor X on the cell surface by a crucial role of the TF transmembrane domain to membrane anchor the reaction, by the TF extracellular domain to provide protein-protein interactions with VIIa to enhance the activity of the catalytic domain of VIIa, and the preferential presentation of factor X as a substrate when associated with phospholipid surfaces.
Ruf
W
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Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Rehemtulla
A
A
Morrissey
J H
JH
Edgington
T S
TS
D000595:Amino Acid Sequence; D001483:Base Sequence; D015942:Factor VIIa / Q000378:metabolism*; D005170:Factor X / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000378:metabolism*; D013379:Substrate Specificity; D013925:Thromboplastin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D010743:Phospholipids; D005170:Factor X; D013925:Thromboplastin; D015942:Factor VIIa