Binding of inactive mutant tissue plasminogen activator to human umbilical vein endothelial cells

1989 ◽  
Vol 3 ◽  
pp. 11
Author(s):  
anitha Ramakrishnan ◽  
Kathleen B. Bechtol ◽  
Walter C. Darbone ◽  
Joffre B. Baker
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Tissue Plasminogen ◽  
Umbilical Vein Endothelial Cells

scholarly journals Thrombin regulation of mRNA levels of tissue plasminogen activator and plasminogen activator inhibitor-1 in cultured human umbilical vein endothelial cells

Blood ◽  
1989 ◽  
Vol 74 (1) ◽  
pp. 222-228 ◽  
Author(s):  
D Dichek ◽  
T Quertermous
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Umbilical Vein ◽  
Plasminogen Activator Inhibitor ◽  
Mrna Levels ◽  
Human Umbilical Vein ◽  
Tissue Plasminogen ◽  
Umbilical Vein Endothelial Cells ◽  
Activator Inhibitor ◽  
Pai 1

Abstract Cultured human umbilical vein endothelial cells release tissue plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) in response to alpha thrombin stimulation. In order to study the mechanisms of thrombin stimulation, we measured changes in levels of mRNA for t-PA and PAI-1 following exposure of endothelial cells to 3 U/mL alpha thrombin. Alpha thrombin causes a significant and time- dependent increase in the mRNA levels of both t-PA and PAI-1. Catalytically inactivated diisofluorophosphate (DIP) treated thrombin and alpha thrombin pretreated with hirudin do not alter t-PA and PAI-1 mRNA levels. We conclude that the increased secretion of t-PA and PAI-1 by human umbilical vein endothelial cells in response to alpha thrombin is mediated at least partially through an increase in mRNA levels. In addition, an active thrombin catalytic site is required for these increases in mRNA to occur.

scholarly journals Thrombin regulation of mRNA levels of tissue plasminogen activator and plasminogen activator inhibitor-1 in cultured human umbilical vein endothelial cells

Blood ◽  
1989 ◽  
Vol 74 (1) ◽  
pp. 222-228 ◽  
Author(s):  
D Dichek ◽  
T Quertermous
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Umbilical Vein ◽  
Plasminogen Activator Inhibitor ◽  
Mrna Levels ◽  
Human Umbilical Vein ◽  
Tissue Plasminogen ◽  
Umbilical Vein Endothelial Cells ◽  
Activator Inhibitor ◽  
Pai 1

Cultured human umbilical vein endothelial cells release tissue plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) in response to alpha thrombin stimulation. In order to study the mechanisms of thrombin stimulation, we measured changes in levels of mRNA for t-PA and PAI-1 following exposure of endothelial cells to 3 U/mL alpha thrombin. Alpha thrombin causes a significant and time- dependent increase in the mRNA levels of both t-PA and PAI-1. Catalytically inactivated diisofluorophosphate (DIP) treated thrombin and alpha thrombin pretreated with hirudin do not alter t-PA and PAI-1 mRNA levels. We conclude that the increased secretion of t-PA and PAI-1 by human umbilical vein endothelial cells in response to alpha thrombin is mediated at least partially through an increase in mRNA levels. In addition, an active thrombin catalytic site is required for these increases in mRNA to occur.

Effect of Lipoprotein(a) and LDL on Plasminogen Binding to Extracellular Matrix and on Matrix-dependent Plasminogen Activation by Tissue Plasminogen Activator

1996 ◽  
Vol 75 (03) ◽  
pp. 497-502 ◽  
Author(s):  
Hadewijch L M Pekelharing ◽  
Henne A Kleinveld ◽  
Pieter F C.C.M Duif ◽  
Bonno N Bouma ◽  
Herman J M van Rijn
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Binding Sites ◽  
Umbilical Vein ◽  
Single Step ◽  
Plasminogen Activation ◽  
Structural Homology ◽  
Tissue Plasminogen ◽  
Umbilical Vein Endothelial Cells

SummaryLp(a) is an LDL-like lipoprotein plus an additional apolipoprotein apo(a). Based on the structural homology of apo(a) with plasminogen, it is hypothesized that Lp(a) interferes with fibrinolysis. Extracellular matrix (ECM) produced by human umbilical vein endothelial cells was used to study the effect of Lp(a) and LDL on plasminogen binding and activation. Both lipoproteins were isolated from the same plasma in a single step. Plasminogen bound to ECM via its lysine binding sites. Lp(a) as well as LDL were capable of competing with plasminogen binding. The degree of inhibition was dependent on the lipoprotein donor as well as the ECM donor. When Lp(a) and LDL obtained from one donor were compared, Lp(a) was always a much more potent competitor. The effect of both lipoproteins on plasminogen binding was reflected in their effect on plasminogen activation. It is speculated that Lp(a) interacts with ECM via its LDL-like lipoprotein moiety as well as via its apo(a) moiety.

Interaction of tissue plasminogen activator with a human endothelial cell 45-kilodalton plasminogen receptor

1994 ◽  
Vol 72 (3-4) ◽  
pp. 126-131 ◽  
Author(s):  
Anil K. Dudani ◽  
Agatha Pluskota ◽  
Peter R. Ganz
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Receptor Binding ◽  
Umbilical Vein ◽  
Annexin Ii ◽  
Cell Protein ◽  
Specific Manner ◽  
Tissue Plasminogen

We have investigated the interaction of tissue plasminogen activator (tPA) with endothelial cell proteins of the human umbilical vein using the technique of ligand blotting. It was observed that tPA interacted with a 45-kilodalton (kDa) endothelial cell protein which appeared to be similar to the 45-kDa plasminogen receptor. Binding of tPA to the 45-kDa protein could be inhibited by excess cold tPA. Morever, excess lysine could inhibit the binding of tPA to the 45-kDa protein in both coincubation and reversibility experiments. These studies indicated that like plasminogen, tPA interacts with the 45-kDa protein in a kringle-dependent and specific manner. To confirm that tPA and plasminogen are interacting with the same protein, we investigated the effect of excess cold plasminogen on tPA binding and excess cold tPA on plasminogen binding in reversibility experiments. It was observed that binding of tPA to the 45-kDa protein was reduced by plasminogen and vice versa. In addition, the 45-kDa protein did not cross-react with antibodies to annexin II, a 40-kDa protein that binds plasminogen and tPA. These latter properties distinguish the 45-kDa receptor from plasminogen/tPA-binding proteins described by others. Therefore, the above studies suggest that the 45-kDa protein represents a unique plasminogen/tPA receptor on human venous endothelial cells.Key words: plasminogen, tissue plasminogen activator, endothelial cells, receptors.

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