title: The Role of Putative Phosphatidylserine-Interactive Residues of Tissue Factor on Its Coagulant Activity at the Cell Surface.
abstract: Exposure of phosphatidylserine (PS) on the outer leaflet of the cell membrane is thought to play a critical role in tissue factor (TF) decryption. Recent molecular dynamics simulation studies suggested that the TF ectodomain may directly interact with PS. To investigate the potential role of TF direct interaction with the cell surface phospholipids on basal TF activity and the enhanced TF activity following the decryption, one or all of the putative PS-interactive residues in the TF ectodomain were mutated and tested for their coagulant activity in cell systems. Out of the 9 selected TF mutants, five of them -TFS160A, TFS161A, TFS162A, TFK165A, and TFD180A- exhibited a similar TF coagulant activity to that of the wild-type TF. The specific activity of three mutants, TFK159A, TFS163A, and TFK166A, was reduced substantially. Mutation of the glycine residue at the position 164 markedly abrogated the TF coagulant activity, resulting in ~90% inhibition. Mutation of all nine lipid binding residues together did not further decrease the activity of TF compared to TFG164A. A similar fold increase in TF activity was observed in wild-type TF and all TF mutants following the treatment of THP-1 cells with either calcium ionomycin or HgCl2, two agents that are commonly used to decrypt TF. Overall, our data show that a few select TF residues that are implicated in interacting with PS contribute to the TF coagulant activity at the cell surface. However, our data also indicate that TF regions outside of the putative lipid binding region may also contribute to PS-dependent decryption of TF.
authors.affiliation: Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United Sates of America.
authors.forename: Shabbir A
authors.initials: SA
authors.lastname: Ansari
authors.affiliation: Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United Sates of America.
authors.forename: Usha R
authors.initials: UR
authors.lastname: Pendurthi
authors.affiliation: Department of Biological Chemistry, Indian Association for the Cultivation of Science, Kolkata, 700032, India.
authors.forename: Prosenjit
authors.initials: P
authors.lastname: Sen
authors.affiliation: Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United Sates of America.
authors.forename: L Vijaya Mohan
authors.initials: LV
authors.lastname: Rao
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D003029:Coagulants / Q000737:chemistry / Q000378:metabolism*; D003412:Cricetulus; D004789:Enzyme Activation; D015942:Factor VIIa / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D006801:Humans; D009000:Monocytes / Q000378:metabolism; D016296:Mutagenesis; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D013925:Thromboplastin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003029:Coagulants; D010718:Phosphatidylserines; D010743:Phospholipids; D011994:Recombinant Proteins; D005170:Factor X; D013925:Thromboplastin; D015942:Factor VIIa