VU Research Portal Modulation of the plasminogen system by thrombin activatable fibrinolysis inhibitor (TAFI) Guimarães, A.H.C. 2006 document version Publisher's PDF, also known as Version of record Link to publication in VU Research Portal citation for published version (APA) Guimarães, A. H. C. (2006). 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E-mail address: [email protected] Download date: 07. Oct. 2021 Modulation of the plasminogen system by thrombin activatable fibrinolysis inhibitor (TAFI) Ana Helena Canas Guimarães ACKNOWLEDGEMENTS Financial support by the Gaubius Laboratory of TNO Prevention and Health in Leiden is gratefully acknowledged. Additional financial support was kindly provided by the J.E. Jurriaanse Stichting and by Kordia Life Sciences. Cover design: Ribbon drawing of the modelled folding of TAFI. Figure adapted from Barbosa Pereira et al. J. Mol. Biol. 2002; 321: 537-47, with permission from the author and from Elsevier, holder of the copyright. Printed by: OPTIMA Grafische Communicatie Rotterdam ISBN 90-8559-217-8 © Ana H.C. Guimarães, 2006 All rights reserved. No part of this book may be reproduced or transmitted in any form or by any means, without prior written permission of the author, or, when appropriate, of the holder of the copyright. VRIJE UNIVERSITEIT Modulation of the plasminogen system by thrombin activatable fibrinolysis inhibitor (TAFI) ACADEMISCH PROEFSCHRIFT ter verkrijging van de graad Doctor aan de Vrije Universiteit Amsterdam, op gezag van de rector magnificus prof.dr. L.M. Bouter, in het openbaar te verdedigen ten overstaan van de promotiecommissie van de faculteit der Geneeskunde op vrijdag 3 november 2006 om 10.45 uur in de aula van de universiteit, De Boelelaan 1105 door Ana Helena Canas Guimarães geboren te Braga, Portugal promotor: prof.dr. V.W.M. van Hinsbergh copromotor: dr. D.C. Rijken The studies presented in this thesis were performed at the Department of Biomedical Research of TNO Prevention and Health, Gaubius Laboratory, Leiden, at the Laboratory of Physiology of the Institute for Cardiovascular Research, Amsterdam and at the Department of Hematology of the Erasmus MC, University Medical Center Rotterdam. “I am a part of all that I have met“ Alfred Lord Tennyson Para o Rao, o meu anjo da guarda. Contents List of abbreviations 8 Introduction Chapter 1 Introduction 12 Biochemical properties of TAFI Chapter 2 Migration of the activation peptide of thrombin activatable 38 fibrinolysis inhibitor (TAFI) during SDS-polyacrylamide gel electrophoresis. (Journal of Thrombosis and Haemostasis 2004; 2: 780-84) Assaying TAFI Chapter 3 Association between thrombin activatable fibrinolysis inhibitor 50 (TAFI) genotype and levels in plasma. Comparison of different assays. (British Journal of Haematology 2004; 124: 659-65) Chapter 4 A new functional assay of thrombin activatable fibrinolysis inhibitor. 66 (Journal of Thrombosis and Haemostasis 2005; 3: 1284-92) Chapter 5 High functional levels of thrombin activatable fibrinolysis inhibitor 86 (TAFI) are associated with an increased risk of first ischemic stroke. (Journal of Thrombosis and Haemostasis 2005; 3: 2211-18) Modulation of the plasminogen system by TAFI Chapter 6 Thrombin activatable fibrinolysis inhibitor (TAFI) affects fibrinolysis 106 in a plasminogen activator concentration-dependent manner. Study of seven plasminogen activators in an internal clot lysis model. (Thrombosis and Haemostasis 2004; 91: 473-79) Chapter 7 Fibrinolytic efficacy of Amediplase, Tenecteplase and scu-PA in 122 different external plasma clot lysis models. Sensitivity for the inhibitory action of thrombin activatable fibrinolysis inhibitor (TAFI). (Thrombosis and Haemostasis 2006; 96: 325-30) Chapter 8 Involvement of thrombin activatable fibrinolysis inhibitor (TAFI) 138 and pancreatic carboxypeptidase B (CPB) in the modulation of capillary tube formation by microvascular endothelial cells. (submitted for publication) Discussion Chapter 9 General discussion 160 Summary / Samenvatting English summary 180 Nederlandse samenvatting 183 Publications 187 Curriculum Vitae 189 Obrigada 191 8 MODULATION OF THE PLASMINOGEN SYSTEM BY TAFI Abbreviations Abbreviations α2-AP α2-antiplasmin LPS lipopolysaccharide, bacterial (see also plasmin inhibitor) mRNA messenger ribonucleic acid aa amino acids MERGETPA DL-2-mercaptomethyl-3-guanidino- APC activated protein C ethylthiopropanoic acid APSAC anisoylated plasminogen-streptokinase MI myocardial infarction activator complex MoAb Monoclonal antibody bFGF basic fibroblast growth factor BSA bovine serum albumin PA plasminogen activator CAD coronary artery disease PAI-1 plasminogen activator inhibitor-1 CABG coronary artery bypass grafting PBS phosphate buffered saline C/EBP CCAAT/enhancer-binding protein PC protein C CPB pancreatic carboxypeptidase B PCI potato carboxypeptidase inhibitor CPN carboxypeptidase N plasma proCPB plasma procarboxypeptidase B CPR carboxypeptidase R PCR polymerase chain reaction CPU carboxypeptidase U PI plasmin inhibitor CRP C-reactive protein Plg plasminogen DNA deoxyribonucleic acid PNP pooled normal plasma DD(E) complex of D-dimer non-convalently associated with fragment E PPACK H-D-Phe-Pro-Arg-chloromethylketone dNTP deoxyribonucleotide triphosphate scu-PA single-chain urokinase-type PA DTT dithiothreitol SNP single nucleotide polymorphism DSPA desmodus rotundus salivary PA SDS-PAGE sodium dodecyl sulphate DVT deep venous thrombosis polyacrylamide gel electrophoresis SD standard deviation εACA ε-amino caproic acid ELISA enzyme-linked immunosorbent assay SEM standard error of the mean EDTA ethylenediaminetetraacetic acid STA staphylokinase TAFI thrombin activatable fibrinolysis FXIII factor XIII inhibitor FCS fetal calf serum TAFIa activated TAFI FbDP fibrin degradation products TAFIai inactivated TAFIa FII prothrombin TBS Tris buffered saline FIIa thrombin TF tissue factor FITC fluorescein isothiocyanate TM thrombomodulin GEMSA guanidinoethyl-mercaptosuccinic acid TNFα tumor necrosis factor α GRE glucocorticoid response element Hepes N-2-hydroxyethylpiperazine-N’-2- TNK-tPA Tenecteplase ethane sulfonic acid tPA tissue-type PA HepG2 human hepatoma cell line tcu-PA two-chain urokinase-type PA hMVEC human microvascular endothelial cell Tris tris(hydroxymethyl)aminoethane HMw high molecular weight HPLC high performance liquid r-Hir recombinant hirudin chromatography r-tPA recombinant tPA HS human serum RT-PCR reverse transcriptase-polymerase IL-1 interleukin 1 chain reaction IL-6 interleukin 6 UAP unstable angina pectoris IS ischaemic stroke uPA urokinase-type plasminogen activator K2tu-PA Amediplase 3’-UTR 3’-untranslated region THESIS MODULATION OF THE PLASMINOGEN SYSTEM BY THROMBIN ACTIVATABLE FIBRINOLYSIS INHIBITOR (TAFI) INTRODUCTION CHAPTER 1 Introduction 12 CHAPTER 1 Introduction Thrombin activatable fibrinolysis inhibitor Thrombin activatable fibrinolysis inhibitor (TAFI, EC 3.4.17.20) was discovered at the end of the 80s, when two groups identified independently an unstable carboxypeptidase present in serum preparations. They named it carboxypeptidase R [1] and carboxypeptidase U [2], respectively. Another group [3] was studying the mechanism of the antifibrinolytic action of thrombin in tissue-type plasminogen activator (tPA)-mediated plasma clot lysis. They spent the following years searching for the “de Fouw Factor”, which was responsible for this effect. Still, this puzzle only started to be unveiled [4] when TAFI was identified as a contaminant during the purification of α2-antiplasmin and named plasma procarboxypeptidase B [5]. Finally in 1995, the protein was rediscovered when seeking to explain the profibrinolytic effect of activated protein C (APC) [6]. The protein was named TAFI because it could be activated by thrombin and when active inhibited fibrinolysis. TAFI has been suggested to circulate in a complex with plasminogen [5,7] but this seems to be based only on its affinity for plasminogen in a purified system and direct evidence is still missing. TAFI was shown to bind to plasminogen, which was immobilised on a Sepharose column and, using a BIAcore system, it was found that the affinity of TAFI for Lys-plasminogen was 10-fold higher than for Glu-plasminogen (Kd - 0.035 µM and Kd - 0.3 µM, respectively) [7]. TAFI is a basic procarboxypeptidase, which is synthesised in the liver and released into the circulation as a single-chain glycoprotein of 60 kDa (Fig.1). Four of the five potential N-linked glycosylation sites are located on the activation peptide with the carbohydrates accounting for about 20% of the zymogen total mass [5,8-10]. As a result, the activation peptide migrates