title: Tissue factor regulates plasminogen binding and activation.
abstract: Tissue factor (TF) has been implicated in several important biologic processes, including fibrin formation, atherogenesis, angiogenesis, and tumor cell migration. In that plasminogen activators have been implicated in the same processes, the potential for interactions between TF and the plasminogen activator system was examined. Plasminogen was found to bind directly to the extracellular domain of TF apoprotein (amino acids 1-219) as determined by optical biosensor interaction analysis. A fragment of plasminogen containing kringles 1 through 3 also bound to TF apoprotein, whereas isolated kringle 4 and miniplasminogen did not. Expression of TF on the surface of a stably transfected Chinese hamster ovary (CHO) cell line stimulated plasminogen binding to the cells by 70% more than to control cells. Plasminogen bound to a site on the TF apoprotein that appears to be distinct from the binding site for factors VII and VIIa as judged by a combination of biosensor and cell assays. TF enhanced two-chain urokinase (tcuPA) activation of Glu-plasminogen, but not of miniplasminogen, in a dose-dependent, saturable manner (half maximal stimulation at 59 pmol/L). TF apoprotein induced an effect similar to that of relipidated TF, but a relatively higher concentration of the apoprotein was required (half maximal stimulation at 3.8 nmol/L). The stimulatory effect of TF on plasminogen activation was confirmed when plasmin formation was examined directly on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In accord with this, TF inhibited fibrinolysis by approximately 74% at a concentration of 14 nmol/L and almost totally inhibited the binding of equimolar concentrations of plasminogen to human umbilical vein endothelial cells and human trophoblasts. Further, CHO cells expressing TF inhibited uPA-mediated fibrinolysis relative to a wild-type control. TF apoprotein and plasminogen were found to colocalize in atherosclerotic plaque. These data suggest that plasminogen localization and activation may be modulated at extravascular sites through a high-affinity interaction between kringles 1 through 3 of plasminogen and the extracellular domain of TF.
authors.affiliation: Department of Pathology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Fan
authors.forename: P J
authors.initials: PJ
authors.lastname: Larson
authors.forename: J
authors.initials: J
authors.lastname: Bognacki
authors.forename: P N
authors.initials: PN
authors.lastname: Raghunath
authors.forename: J E
authors.initials: JE
authors.lastname: Tomaszewski
authors.forename: A
authors.initials: A
authors.lastname: Kuo
authors.forename: G
authors.initials: G
authors.lastname: Canziani
authors.forename: I
authors.initials: I
authors.lastname: Chaiken
authors.forename: D B
authors.initials: DB
authors.lastname: Cines
authors.forename: A A
authors.initials: AA
authors.lastname: Higazi
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D015374:Biosensing Techniques; D016466:CHO Cells; D002478:Cells, Cultured; D003324:Coronary Artery Disease / Q000378:metabolism / Q000473:pathology; D006224:Cricetinae; D003412:Cricetulus; D004730:Endothelium, Vascular / Q000166:cytology; D005167:Factor VII / Q000302:isolation & purification / Q000378:metabolism; D005341:Fibrinolysin / Q000096:biosynthesis; D005342:Fibrinolysis / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D018082:Kringles / Q000502:physiology; D010743:Phospholipids / Q000378:metabolism; D010958:Plasminogen / Q000737:chemistry / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology
chemical_list: D001059:Apoproteins; D010743:Phospholipids; D011993:Recombinant Fusion Proteins; D005167:Factor VII; D010958:Plasminogen; D013925:Thromboplastin; D010960:Plasminogen Activators; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator