Plasminogen: Structure,Activation, and Regulation

Plasminogen: Structure, Activation, and Regulation

Edited by

DAVID M. WAISMAN Department of Biochemistry & Molecular Biology and Oncology University of Calgary Health Sciences Centre Calgary, Alberta, Canada

Springer Science+Business Media, LLC

Library of Congress Cataloging-in-Publication Data

Plasminogen: structure, activation, and regulation/edited by David Morton Waisman. p. ; cm.

Includes bibliographical references and index. ISBN 978-1-4613-4949-5 ISBN 978-1-4615-0165-7 (eBook) DOI 10.1007/978-1-4615-0165-7

1. Plasminogen. 1. Waisman, David Morton, 1952-

QP93.5 .W356 2003 612'.0151-dc21

ISBN 978-1-4613-4949-5

©2003 Springer Science+Business Media New York Originally published by Kluwer Academic / Plenum Publishers in 2003 Softcover reprint of the hardcover Ist edition 2003

AII rights reserved

2002042762

No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher, with the exception of any material supplied specifically for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work.

To my wife, Mary Sue, and children Meredith and Phillipfor their unfailing support and patience

David M. Waisman

Contributors

Riitta AlitaloDepartment of Clinical ChemistryHematology and Stem Cell Laboratories andHelsinki University Central Hospital

Finland

Elizabeth H. AllanSt Vincent's Institute of Medical Research9 Princes StreetFitzroy Vic 3065Australia

Nicholas M. AndronicosThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Toomas AsserDepartment of NeurologyUniversity of TartuTartu, Estonia

Felizabel Garcia BannachThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Francis J. CastellinoW. M. Keck Center for Transgene ResearchDepartment of Chemistry and BiochemistryUniversity of Notre DameNotre Dame, Indiana 46556United States

Kyu-Sil ChoiDepanment of Biochemistry and Molecular BiologyUniversity of CalgaryCalgary, AlbenaCanada TIN 4Nl

Desire CollenCenter for Molecular and Vascular BiologyUniversity of LeuvenCampus Gasthuisberg0& N, Herestraat 49B-3000 LeuvenBelgium

vii

Evis DaciOnderwijs en NavorsingGasthuisberg3000 LeuvenBelgium

Paul J. DeclerckLaboratory for Pharmaceutical BiologyFaculty of Pharmaceutical SciencesKatholieke Universiteit LeuvenVan Evenstraat 4B-3000 LeuvenBelgium

Yves A. DeClerckDivision of Hematology-Oncology andDepanments of Pediatrics and Biochemistry andMolecular Biology

Childrens Hospital Los Angeles and USC KeckSchool of Medicine

Los Angeles, CaliforniaUnited States

Vincent EllisSchool of Biological SciencesUniversity of East AngliaNorwich NR4 7TJUnited Kingdom

Jordi FelezInstitut de Recerca Oncologica08907, BarcelonaSpain

Darin K. FoggDepartment of Biochemistry and Molecular BiologyUniversity of CalgaryCalgary, AlbenaCanada TIN 4N I

Ann GilsLaboratory for Pharmaceutical BiologyFaculty of Pharmaceutical SciencesKatholieke Universiteit LeuvenVan Evenstraat 4B-3000 LeuvenBelgium

viii

Neill A. GinglesThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Yun GongThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Davida K. GrellaUniversity of Notre DameNotre Dame, Indiana

Ana Gutierrez-FernandezThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Philip J. HoggCentre for Thrombosis and Vascular ResearchSchool of Medical SciencesUniversity of New South Wales and Department ofHaematology

Prince of Wales HospitalSydney, NSW 2052Australia

Jane L. Hoover-PlowJoseph 1. Jacobs Center for Thrombosis andVascular Biology

Department of Molecular CardiologyLerner Research InstituteCleveland Clinic FoundationCleveland, Ohio 44195United States

G. Ronald JenkinsThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Andres KullaDepartment of Pathology and NeuropathologyTartu University ClinicsTartu, Estonia

Contributors

Mijung KwonCancer Biology Research GroupDepartment of Biochemistry and Molecular BiologyUniversity of CalgaryCalgary, AlbertaCanada TIN 4NI

Walter E. LaugDivision of Hematology-Oncology andDepartments of Pediatrics and Biochemistry andMolecular Biology

Childrens Hospital Los Angeles and USC KeckSchool of Medicine

Los Angeles, CaliforniaUnited States

H. Roger LijnenCenter for Molecular and Vascular BiologyUniversity of LeuvenCampus Gasthuisbergo & N, Herestraat 49B-30oo LeuvenBelgium

David J. LoskutoffThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

T. John MartinSt Vincent's Institute of Medical Research9 Princes StreetFitzroy Vic 3065Australia

Lindsey A. MilesThe Scripps Research InstituteDepartment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Satu MustjokiDepartment of Clinical ChemistryHematology and Stem Cell Laboratories andBiomedicum Helsinki

Hematopoietic Stem Cell ResearchHelsinki University Central HospitalFinland

Robert J. ParmerDepartment of MedicineUniversity of California and Veterans AdministrationMedical Center

San Diego, California 92161United States

Contributors

Michael S. PepperDepartment of MorphologyUniversity Medical CenterGeneva, Switzerland

Victoria A. PloplisW. M. Keck Center for Transgene ResearchDepartment of Chemistry and BiochemistryUniversity of Notre DameNotre Dame, Indiana 46556United States

Aadu SimiskerHaartman InstituteUniversity of HelsinkiPOB 21, F1N-00014 HelsinkiFinland

Department of NeurologyUniversity of TartuTartu, Estonia

Tambet TeesaluHaartman Institute,University of HelsinkiPOB 21, FIN-OOOI4 HelsinkiFinland

Department of Pathology and NeuropathologyTanu University clinicsTartu, Estonia

Depanment of NeurologyUniversity of TanuTartu, Estonia

ix

Antti VaheriHaartman Institute,University of HelsinkiPOB 21, F1N-OOOI4 HelsinkiFinland

David M. WaismanCancer Biology Research GroupDepartment of Biochemistry and Molecular BiologyUniversity of CalgaryCalgary, AlbenaCanada T2N 4N I

Lu ZhangThe Scripps Research InstituteDepanment of Cell BiologyDivision of Vascular BiologyLa Jolla, California 92037United States

Preface

The observation that a blood clot spontaneously dissolves was first described by Denysin 1889. Subsequently, the blood clotting system was shown to be involved in tumorgrowth. For example, as early as 1925, Fisher reported that avian tissue explants trans­formed to malignancy by viruses generated high levels of fibrinolytic activity under con­ditions in which cultures of normal cells did not. In 1958, the concept that anequilibrium existed between the tendency of blood to clot and to remain fluid was pro­posed by Astrup. At that time, it was believed that this hemostatic balance was explainedby the ability of polymerizing fibrin to orchestrate its own clearance by stimulating fib­rinolytic activity. Since these pioneering studies, considerable information has accumu­lated that has defined the components of the coagulation and fibrinolytic systems andhow they are involved in physiological and pathophysiological processes. Plasminogen:Structure, activation, and regulation focuses on the basic principles and recent develop­ments in the plasminogen/plasmin research field and how these results provide a con­ceptual framework for an understanding of the physiological role of plasminogen inhealth and disease.

The enzymatic cascade triggered by activation of plasminogen has been implicatedin a variety of normal and pathological events such as fibrinolysis, wound healing, tis­sue remodeling, embryogenesis, angiogenesis, and the invasion and metastasis of tumorcells. This impressive list of physiological functions for plasminogen reinforces the widediversity of roles that plasminogen plays in various physiological processes. Productiveplasmin generation requires the assembly of both plasminogen activators and plasmino­gen on a solid support such as the fibrin polymer or the cell surface. The regulation ofplasmin production involves a complex interplay between these plasminogen activators,plasminogen activator inhibitors, and plasmin inhibitors. Clearly, the explosive growthin this research field and the many exciting discoveries suggests that the research effortsin the next decade will reveal the mechanisms by which the components of the plas­minogen system interact and regulate both plasmin activation and function at a cellularlevel.

Plasminogen: Structure, activation, and regulation is divided into two sections.The first section deals with the structure and regulation of plasminogen. The chaptersin this section range from discussions of the structure of plasminogen and the regulationof the plasminogen gene to discussions of the structure and regulation of plasminogenactivators and plasminogen activator inhibitors. Also examined is the relatively new dataconcerning the generation of anti-angiogenic molecules from plasminogen. The secondsection deals with the physiological and pathophysiological roles of plasminogen as wellas the consequences of plasminogen gene knockout. Discussions in this section includeexamination of the role of plasminogen in hematopoietic malignancies, tumor cellprogression, angiogenesis, mammary gland involution, wound healing, and bonereadsorption.

xi

xii Preface

In closing, I would like to thank my administrative assistant, Ms. Vi Sommerfeld, forher invaluable assistance and timeless efforts with the organization and editing of the book.Lastly, I would like to acknowledge the efforts of the authors of the individual chapters,who are authorities in this field, for agreeing to take time from busy schedules to providethese chapters in a timely fashion.

David Morton Waisman

Contents

Part I. Plasminogen: Structure and Regulation

1. Human Plasminogen: Structure, Activation, and Function

Francis J. Castellino and Victoria A. Ploplis

1. Introduction 32. Structure of Human Plasminogen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32.1. Primary Protein Structure .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32.2. Gene Organization 5

3. Activation of Human Plasminogen . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 63.1. Activation by Physiological Activators 73.1.1. Urokinase-type Plasminogen Activator. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73.1.2. Tissue-type Plasminogen Activator. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

3.2. Activation by Bacterial-derived Plasminogen Activators . . . . . . . . . . . . . . . . . . . . . 93.2.1. Streptokinase 93.2.2. Staphylokinase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

4. Targets for Plasmin Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95. Dysplasminogenemias and Phenotypic Manifestations 106. Conclusions 11References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

2. Plasminogen Activators: Structure and Function

Vincent Ellis

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192. Serine Proteases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203. Urokinase Plasminogen Activator, uPA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213.1. Serine Protease Domain 223.2. N-terminal Domains. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243.2.1. KR Module 243.2.2. EG Module 24

4. Mechanisms Regulating uPA Function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254.1. Zymogen Activation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254.2. Zymogen Activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264.3. Reciprocal Zymogen Activation 274.4. uPAR Stimulation of Plasminogen Activation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274.4.1. uPA and the Template Mechanism 284.4.2. Plasminogen and the Template Mechanism 29

4.5. Avian uPA, a Special Case? 30

xiii

xiv Contents

5. Tissue Plasminogen Activator, tPA .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305.1. Serine Protease Domain 315.2. N-terminal Domains , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

5.2.1. KR Modules , .. , .. , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 335.2.2. F1-EG Supermodule 33

6. Mechanisms Regulating tPA Function 346.1. Zymogenicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 346.2. Fibrin Stimulation of Plasminogen Activation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

6.2.1. tPA/fibrin Interaction 366.2.2. Vampire Bat Plasminogen Activator , , . . . . 37

6.3. Cellular Mechanisms Regulating tPA Activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 376.3.1. Endothelial Cells ,..................................... 386.3.2. Vascular Smooth Muscle Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 386.3.3. Prion Protein ,................................. 38

7. Concluding Remarks ,................................. 39References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

3. Plasminogen Activators Inhibitors

Ann Gils and Paul J. Declerck

1. Plasminogen Activator Inhibitor-1 471.1. Natural and Recombinant PAI-1 ,... 481.2. Distinct Conformations of PAI-1 491.3. Target Specificity of PAI-1 ,....................... 501.4. PAI-1 in Patho-Physiological Processes 51

1.4.1. PAI-1 and Cardiovascular Disease 511.4.2. PAI-1 and Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

1.5. PAI-1 Inhibitors and their Binding Sites , . . . 532. Plasminogen Activator Inhibitor-2 542.1. Function of PAI-2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 542.2. PAI-2 Polymerization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

3. Plasminogen Activator Inhibitor-3 563.1. Target Specificity of PAI-3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 563.2. Physiological Role of PAI-3 56References .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57

4. Regulation of Plasminogen Gene Expression

Ana Gutierrez-Fernandez, Neill A. Gingles, Lu Zhang, Felizabel Garcia Bannach,G. Ronald Jenkins, David J. Loskutoff, Robert J. Parmer, and Lindsey A. Miles

1. Introduction 672. The Plasminogen Gene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 682.1. Mechanisms of Constitutive Regulation of Plasminogen Gene Expression 682.2. Plasminogen Distribution in Tissues , . . . . . . . . . . . . . . . . . . . 68

3. Regulation of Plasminogen Gene Expression in vitro and in vivo 723.1. Interleukin-6 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 723.2. Glucocorticoids 74

Contents xv

3.3. LPS 753.4. Tumor Necrosis Factor-a (TNF-a) and Transforming Growth

Factor-13 (TGF-I3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 763.5. Interleukin-l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 763.6. Kainic Acid 77

4. Conclusions 77Acknowledgments 78References 78

5. Plasminogen Receptors

Darin K. Fogg, Kyu-Sil Choi, and David M. Waisman

1. Introduction to Cellular Plasminogen Activation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 812. Mechanism of Cellular Plasminogen Binding. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 822.1. History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 822.2. Kinetics of Plasminogen Binding 822.3. Binding of Plasminogen Isoforms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

3. Plasminogen Receptors as Regulators of Plasmin Activity 854. Modulation of Receptor Expression 864.1. Plasminogen Receptor Expression on Platelets .. . . . . . . . . . . . . . . . . . . . . . . . . . . 864.2. Plasminogen Receptor Expression on Endothelial Cells 874.3. Plasminogen Receptor Expression on Peripheral Blood Cells 874.4. Plasminogen Receptor Expression on Tumor Cells . . . . . . . . . . . . . . . . . . . . . . . . . 88

5. Candidate Plasminogen Receptors 886. Annexin II, Pll, and Annexin II Heterotetramer as Candidate PlasminogenReceptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91

7. Concluding Remarks 97References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98

6. The Role of Lys-Plasminogen in Cell-Mediated Plasmin Production

Lindsey A. Miles, Nicholas M. Andronicos, Jordi Felez, Davida K. Grella, Francis J.Castellino, and Yun Gong

1. Introduction 1032. Key Differences between Glu-Plasminogen and Lys-Plasminogen 1043. Mechanisms by which Plasminogen Activation is Enhanced on theCell Surface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1063.1. Role of the Glu-Plasminogen to Lys-Plasminogen Conversion in Plasminogen

Activation by t-PA on the Cell Surface 1073.2. Role of the Glu-Plasminogen to Lys-Plasminogen Conversion in Plasminogen

Activation by u-PA on the Cell Surface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1123.3. Effect of Cells on Conversion of [D(646)E]Glu-Pg to [D(646)E]Lys-Pg byExogenous Plasmin 114

4. Conclusions 115Acknowledgments 116References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

xvi

7. Plasmin Reductase

Philip J. Hogg

Contents

1. Introduction 1212. Identification of Plasmin Reductase Activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1223. Disulfide-Bonds Cleaved by Plasmin Reductase. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1244. Proteolysis of Reduced Plasmin 1255. Plasmin Reductase is Phosphoglycerate Kinase 1266. Structure/Function Aspects of Plasmin Reduction by Phosphoglycerate Kinase 1267. Putative Mechanism of Action of Phosphoglycerate Kinase 1288. Phosphoglycerate Kinase in Tumor Angiogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1299. Future Directions 131References ....................•..................................... 131

8. Mechanism of Angiostatin Formation from Plasminogen

Mijung Kwon and David M. Waisman

1. Introduction 1352. Identification of Plasminogen Fragment, AM' Produced by Plasmin Autoproteolysis . . . 1363. Characterization of AM' an Anti-Angiogenic Plasminogen Fragment 1394. Identification of Cell-Generated Plasminogen Fragments 1405. Characterization of p22, the Smallest Anti-Angiogenic Plasminogen FragmentProduced by Cultured Cells 141

6. Identification of Plasminogen Fragments Present in Sera 1437. Annexin II Tetramer, a Putative Plasminogen Receptor. . . . . . . . . . . . . . . . . . . . . . . . . 1438. Plasmin Reductase Activity of Annexin II Tetramer 1469. Mechanism of Action ofAnnexin II Tetramer on A61 Formation. . . . . . . . . . . . . . . . . . 14810. Concluding Remarks 151Acknowledgment 151References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151

Part II. Physiological Roles of Plasminogen

9. Lessons Learned from the Pig Deficient Mice (Pig - / -)

Jane L. Hoover-Plow

1. Introduction 1592. Development 1592.1. Physical Development 1602.2. Behavioral Development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161

3. Role of the PIg System in Stress. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1633.1. Alterations of the PIg System in Response to Stress 1633.2. Grooming, a Stress-Induced Behavior is Increased in the Plg-j- Mice. . . . . . . . 164

Contents xvii

3.3. Acoustic Startle Reflex, a Stress-Induced Behavior is Decreased inPlg-/- Mice. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165

3.4. The PIg System in Neuroendocrine Tissues and Alterations in Plg-/- Mice ..... 1654. Role of the PIg System in Adipose Tissue Development. . . . . . . . . . . . . . . . . . . . . . . . . 1664.1. Role of the PIg System in Obesity 1664.2. PIg and PIg System Components in Adipose Tissue 1674.3. Adipose Tissue Development in Plg-/- Mice. . . . . . . . . . . . . . . . . . . . . . . . . . . . 1684.4. Vascularization of Adipose Tissue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

5. Summary 171References 171

10. Plasminogen-Directed Phenotypes in Mice

Victoria A. Ploplis and Francis J. Castellino

1. Introduction 1772. Generation and Initial Characterization of Plasminogen-DeficientMice (PG-/-) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177

3. Challenge-Induced Phenotypes in Mice Deficient for Plasminogen. . . . . . . . . . . . . . . . . 1783.1. Pulmonary Clot Lysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1783.2. Responses to Vascular Injury 1793.3. Pathogen Susceptibility and Inflammation 1793.4. Glomerulonephritis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1813.5. Rheumatoid Arthritis 1813.6. Pulmonary Fibrosis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1823.7. Wound Healing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1823.8. Neuronal and Axonal Degeneration and Demyelination 1833.9. Tumor Development, Metastatis, and Angiogenesis 183

4. Conclusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184References 184

11. Role of the Plasminogen and MMP Systems in Wound Healing

H. Roger Lijnen and Desire Col/en

1. Introduction 1892. Plasminogen/Plasmin and MMP Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1892.1. PlasminogenlPlasmin System .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1892.2. MMP System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . 1912.3. Molecular Interactions Between the Plasminogen/Plasmin and

MMP Systems 1913. Arterial Restenosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1923.1. Role of the PlasminogenlPlasmin System in Neointima Formation . . . . . . . . . . . . . 1933.2. Role of the MMP System in Neointima Formation . . . . . . . . . . . . . . . . . . . . . . . . . 194

4. Allograft Transplant Stenosis 1955. Skin Wound Healing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1956. Myocardial Ischemia 1967. Summary 196References 196

xviii Contents

12. Matrix Metalloproteinases and the Plasminogen System in TumorProgression

Yves A. DeClerck and Walter E. Laug

1. Introduction 2012. Matrix Metalloproteinases 2022.1. MMP Classification and their Expression in Cancer . . . . . . . . . . . . . . . . . . . . . . . . 2022.2. Substrate Specificity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2042.3. Activation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204

3. Interactions Between MMPs and the Plasminogen System. . . . . . . . . . . . . . . . . . . . . . . 2053.1. Activation of proMMPs by Plasmin 2053.2. Cleavage of Plasmin by MMPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2063.3. MMPs Degrade Plasmin Inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2063.4. MMPs and Plasmin Degrade Several Common Proteins. . . . . . . . . . . . . . . . . . . . . 206

4. Interaction between MMPs and the Plasminogen System in Tumor Invasion andMetastasis 2074.1. Expression of MMPs and PA in Cancer 2074.2. MMPs and Plasminogen Interactions in vitro 2074.3. MMPs and Plasminogen Interaction in vivo 2084.4. Conclusion and Directions 209

5. Interaction between MMPs and Plasminogen System in Angiogenesis 2095.1. In vitro and in vivo Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2095.2. Paradoxical Aspects 210

6. Relevance of these Studies in Human Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211Acknowledgments 211References 212

13. Role of Plasminogen Activation in Hematopoietic Malignancies and inNormal Hematopoiesis

Satu Mustjoki, Riitta Alitalo, and Antti Vaheri

1. Introduction 2172. Normal Hematopoiesis and its Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2173. Leukemia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2183.1. Classification 2183.2. Clinical Findings 219

4. Plasminogen Activation System in General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2204.1. Structure and General Function of uPAR 2214.2. Soluble uPAR (suPAR) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

5. Plasminogen Activation in Leukemia 2235.1. Components of the PA System in Normal Blood Cells 2235.2. Components of the PA System in Leukemia Cells. . . . . . . . . . . . . . . . . . . . . . . . . . 2245.3. Soluble uPA and uPAR in Leukemia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2255.4. Expression Patterns of uPAR Fragments in Leukemia Patients

and in Healthy Controls 2266. Clinical Impact of Plasminogen Activation in Leukemia . . . . . . . . . . . . . . . . . . . . . . . . . 2276.1. Plasminogen Activation System and Hemostasis . . . . . . . . . . . . . . . . . . . . . . . . . . . 2276.2. Cell Surface Plasminogen Activators and Urokinase Receptor: Diagnostic and

Prognostic Tools in Leukemia? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2286.3. suPAR as a Marker for Prognosis in Leukemia . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

Contents xix

7. Perspectives 230References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230

14. The Role of Plasminogen in Bone Remodeling

Evis Daci, Elizabeth H. Allan, and T. John Martin

1. Osteoblasts and Bone Formation 2372. Osteoclasts and Bone Resorption 2393. Bone Remodeling. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2404. Regulation of the PAlPlasmin System in Bone Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2414.1. Peptide Hormones and Cyclic AMP 2414.2. Glucocorticoids and 1,25(OH)z Vitamin D3 ..........••.........••.....•. 2424.3. Cytokines and Growth Factors 242

5. Potential Roles of the PA/plasmin System in Bone. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2435.1. Bone Resorption. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2435.2. Studies in Genetically Manipulated Mice 2445.3. Bone Formation 246References .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246

15. Plasminogen Activators in eNS Physiology and Disease

Tambet Teesalu, Andres Kulla, Toomas Asset; Aadu Simisket; andAntti Vaheri

1. Introduction 2512. Expression and Function of Plasminogen Activators in the Normal CNS 2522.1. Plasminogen Activators in Developing and Adult CNS ... . . . . . . . . . . . . . . . . . . . 2522.2. Role of Plasminogen Activators in Neuronal Migration, Plasticity, and Learning .. 2542.2.1. Neuronal Migration 2552.2.2. Synaptic Plasticity 255

3. Plasminogen Activators in CNS Pathologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2563.1. Excitotoxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2563.2. Ischemic Stroke 2573.3. Neurodegenerative Diseases 2593.3.1. Alzheimer's Disease. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2593.3.2. Creutzfeldt-Jakob Disease 260

3.4. Neuroinflammatory Diseases 2603.5. CNS Malignancy 261

4. Conclusions and Future Perspectives 2635. Acknowledgments 263References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263

16. Role of the Plasminogen Activator-Plasmin System inAngiogenesis

Michael S. Pepper

1. Introduction 2692. Role of the PA-Plasmin System in Angiogenesis 2732.1. uPA/uPAR Interactions are Required for Angiogenesis. . . . . . . . . . . . . . . . . . . . . . 275

xx Contents

2.2. PAI-I is Required for Angiogenesis 2762.3. Does tPA Playa Role in Angiogenesis? 2772.4. MMP Requirement for Fibrinolysis During Angiogenesis . . . . . . . . . . . . . . . . . . . . 2782.5. Angiostatin and other Angiostatic Derivatives of Plasminogen 279

3. Requirement for the PA-Plasmin Systems in Polyoma Virus MiddleT Oncogene-induced Vascular Tumor Fonnation 280

4. Conclusions and Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 282Acknowledgments 283References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283

Index 291

Plasminogen: Structure,Activation, and Regulation