Structural Mechanism for Heparin-Binding of the Third Kunitz Domain of Human Tissue Factor Pathway Inhibitor†,‡

Biochemistry ◽  
2002 ◽  
Vol 41 (1) ◽  
pp. 78-85 ◽  
Author(s):  
Shouhei Mine ◽  
Toshio Yamazaki ◽  
Toshiyuki Miyata ◽  
Saburo Hara ◽  
Hisao Kato
Keyword(s):  
Tissue Factor ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Heparin Binding ◽  
Structural Mechanism ◽  
The Third ◽  
Kunitz Domain ◽  
Tissue Factor Pathway

scholarly journals Structural and functional characterization of tissue factor pathway inhibitor following degradation by matrix metalloproteinase-8

2002 ◽  
Vol 367 (2) ◽  
pp. 451-458 ◽  
Author(s):  
Anna C. CUNNINGHAM ◽  
Karen A. HASTY ◽  
Jan J. ENGHILD ◽  
Alan E. MAST
Keyword(s):  
Matrix Metalloproteinase ◽  
Tissue Factor ◽  
Tissue Factor Pathway Inhibitor ◽  
Proteolytic Enzymes ◽  
Functional Characterization ◽  
Factor Xa ◽  
Terminal Region ◽  
The Third ◽  
Kunitz Domain ◽  
Tissue Factor Pathway

Vascular injury results in the activation of coagulation and the release of proteolytic enzymes from neutrophils and connective- tissue cells. High concentrations of these inflammatory proteinases may destroy blood coagulation proteins, contributing to coagulation and bleeding disorders associated with severe inflammation. Matrix metalloproteinase-8 (MMP-8) is released from neutrophils at sites of inflammation and vascular disease. We have investigated the effect of MMP-8 degradation on the anticoagulant function of tissue factor pathway inhibitor (TFPI) as a potential pathological mechanism contributing to coagulation disorders. MMP-8 cleaves TFPI following Ser174 within the connecting region between the second and third Kunitz domains (kcat/Km75M-1·s-1) as well as following Lys20 within the NH2-terminal region. MMP-8 cleavage of TFPI decreases the anticoagulant activity of TFPI in factor Xa initiated clotting assays as well as the ability of TFPI to inhibit factor Xa in amidolytic assays. Yet, MMP-8 cleavage does not alter the ability of TFPI to inhibit trypsin. Since the inhibition of both factor Xa and trypsin is mediated by binding to the second Kunitz domain, these results suggest that regions of TFPI other than the second Kunitz domain may directly interact with factor Xa. 125I-factor Xa ligand blots of TFPI fragments generated following MMP-8 degradation were used for probing binding interactions between factor Xa and regions of TFPI, other than the second Kunitz domain. In experiments performed under reducing conditions that disrupt the Kunitz domain structure, 125I-factor Xa binds to the C-terminal fragment of MMP-8-degraded TFPI. This fragment contains portions of TFPI distal to Ser174, which include the third Kunitz domain and the basic C-terminal region. An altered form of TFPI lacking the third Kunitz domain, but containing the C-terminal region, was used to demonstrate that the C-terminal region directly interacts with factor Xa.

The second Kunitz domain of human tissue factor pathway inhibitor: cloning, structure determination and interaction with factor Xa 1 1Edited by T. Richmond

1997 ◽  
Vol 269 (3) ◽  
pp. 395-407 ◽  
Author(s):  
Maurits J.M Burgering ◽  
Leon P.M Orbons ◽  
Antoon van der Doelen ◽  
John Mulders ◽  
Henri J.M Theunissen ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Structure Determination ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Factor Xa ◽  
Kunitz Domain ◽  
Tissue Factor Pathway

scholarly journals Engineering Kunitz Domain 1 (KD1) of Human Tissue Factor Pathway Inhibitor-2 to Selectively Inhibit Fibrinolysis

2010 ◽  
Vol 286 (6) ◽  
pp. 4329-4340 ◽  
Author(s):  
Madhu S. Bajaj ◽  
Godwin I. Ogueli ◽  
Yogesh Kumar ◽  
Kanagasabai Vadivel ◽  
Gregory Lawson ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Kunitz Domain ◽  
Tissue Factor Pathway

Pharmacokinetics of Full Length and Two-Domain Tissue Factor Pathway Inhibitor in Combination with Heparin in Rabbits

1993 ◽  
Vol 70 (03) ◽  
pp. 454-457 ◽  
Author(s):  
Claus Bregengaard ◽  
Ole Nordfang ◽  
Per Østergaard ◽  
Jens G L Petersen ◽  
Giorgio Meyn ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Tissue Factor Pathway Inhibitor ◽  
Anticoagulant Activity ◽  
Fold Increase ◽  
Volume Of Distribution ◽  
Full Length ◽  
Heparin Binding ◽  
Extrinsic Pathway ◽  
C Terminus ◽  
Tissue Factor Pathway

SummaryTissue factor pathway inhibitor (TFPI) is a feed back inhibitor of the initial activation of the extrinsic pathway of coagulation. In humans, injection of heparin results in a 2-6 fold increase in plasma TFPI and recent studies suggest that TFPI may be important for the anticoagulant activity of heparin. Full length (FL) TFPI, but not recombinant two-domain (2D) TFPI, has a poly cationic C-terminus showing very strong heparin binding. Therefore, we have investigated if heparin affects the pharmacokinetics of TFPI with and without this C-terminus.FL-TFPI (608 U/kg) and 2D-TFPI (337 U/kg) were injected intravenously in rabbits with and without simultaneous intravenous injections of low molecular weight heparin (450 anti-XaU/kg).Heparin decreased the volume of distribution and the clearance of FL-TFPI by a factor 10-15, whereas the pharmacokinetics of 2D-TFPI were unaffected by heparin. When heparin was administered 2 h following TFPI the recovery of FL-TFPI was similar to that found in the group receiving the two compounds simultaneously, suggesting that the releasable pool of FL-TFPI is removed very slowly in the absence of circulating heparin.

Factor Xa Enhances the Binding of Tissue Factor Pathway Inhibitor to Acidic Phospholipids

1996 ◽  
Vol 75 (05) ◽  
pp. 796-800 ◽  
Author(s):  
Sanne Valentin ◽  
Inger Schousboe
Keyword(s):  
Tissue Factor ◽  
Tissue Factor Pathway Inhibitor ◽  
Factor Xa ◽  
Microtitre Plate ◽  
C Terminus ◽  
Microtitre Plate Assay ◽  
Kunitz Domain ◽  
Tissue Factor Pathway ◽  
Acidic Phospholipids

SummaryIn the present study, the interaction between tissue factor pathway inhibitor (TFPI) and phospholipids has been characterized using a microtitre plate assay. TFPI was shown to bind calcium-independently to an acidic phospholipid surface composed of phosphatidylserine, but not a surface composed of the neutral phosphatidylcholine. The interaction was demonstrated to be dependent on the presence of the TFPI C-terminus. The presence of heparin (1 U/ml, unfractionated) was able to significantly reduce the binding of TFPI to phospholipid. The interaction of TFPI with phosphatidylserine was significantly decreased in the presence of calcium, but this was counteracted, and even enhanced, following complex formation of TFPI with factor Xa prior to incubation with the phospholipid surface. Moreover, a TFPI variant, not containing the third Kunitz domain and the C-terminus, was unable to bind to phospholipid. However, following the formation of a TFPI/factor Xa-complex this TFPI variant was capable of interacting with the phospholipid surface. This indicates that the role of factor Xa as a TFPI cofactor, at least in part, is to mediate the binding of TFPI to the phospholipid surface.

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