Queen's University - Utility Bar

QSpace at Queen's University >
Theses, Dissertations & Graduate Projects >
Queen's Theses & Dissertations >

Please use this identifier to cite or link to this item: http://hdl.handle.net/1974/6070

Title: A Quantitative and Mechanistic Assessment of Activated Thrombin-Activatable Fibrinolysis Inhibitor and its Role in Pathological Bleeding and Thrombosis
Authors: Foley, Jonathan

Files in This Item:

File Description SizeFormat
Foley_Jonathan_H_201009_PhD.pdf822.74 kBAdobe PDFView/Open
Keywords: enzymology
carboxypeptidase
hemophilia
fibinrolysis
Issue Date: 2010
Series/Report no.: Canadian theses
Abstract: The coagulation and fibrinolytic systems are linked by the thrombin-thrombomodulin complex which regulates each system through activation of protein C and TAFI, respectively. We have used novel assays and techniques to study the enzymology and biochemistry of TAFI and TAFIa, to measure TAFI activation in hemophilia A and protein C deficiency and to determine if enhancing TAFI activation can improve hemostasis in hemophilic plasma and whole blood. We show that TAFIa not TAFI attenuates fibrinolysis in vitro and this is supported by a relatively high catalytic efficiency (16.41μM-1s-1) of plasminogen binding site removal from fibrin degradation products (FDPs) by TAFIa. Since the catalytic efficiency of TAFIa in removing these sites is ~60-fold higher than that for inflammatory mediators such as bradykinin it is likely that FDPs are a physiological substrate of TAFIa. The high catalytic efficiency is primarily a result of a low Km which can be explained by a novel mechanism where TAFIa forms a binary complex with plasminogen and is recruited to the surface of FDPs. The low Km also suggests that TAFIa would effectively cleave lysines from FDPs during the early stages of fibrinolysis (i.e. at low concentrations of FDPs). Since individuals with hemophilia suffer from premature fibrinolysis as a result of insufficient TAFI activation we quantified TAFI activation in whole blood from hemophilic subjects. Both the rate of activation and the area under the TAFI activation time course (termed TAFIa potential) was determined to be reduced in hemophilia A and the TAFIa potential was significantly and inversely correlated with the clinical bleeding iii phenotype. Using a novel therapeutic strategy, we used soluble thrombomodulin to increase TAFI activation which improved the clot lysis time in factor VIII deficient human plasma and hemophilic dog plasma as well as hemophilic dog blood. Finally, we briefly show in a biochemical case study that TAFI activation is enhanced in protein C deficiency and when afflicted individuals are placed on Warfarin anticoagulant therapy, TAFI activation is reduced. Since TAFIa stabilizes blood clots, this suggests that reducing TAFI activation or inhibiting TAFIa may help restore blood flow in vessels with pathological thrombosis.
Description: Thesis (Ph.D, Biochemistry) -- Queen's University, 2010-09-23 10:12:18.892
URI: http://hdl.handle.net/1974/6070
Appears in Collections:Biochemistry Graduate Theses
Queen's Theses & Dissertations

Items in QSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

 

  DSpace Software Copyright © 2002-2008  The DSpace Foundation - TOP