| Structure | |
| X-Ray Crystal Structures of Human [alpha]-Thrombin and of the Human Thrombin-Hirudin Complex | p. 3 |
| Thrombin Structure and Function | p. 3 |
| Crystal Structure Analysis of PPACK-Human [alpha]-Thrombin | p. 6 |
| Overall Structure of Human [alpha]-Thrombin | p. 7 |
| Comparison with Related Serine Proteinase Structures | p. 13 |
| The Role of Charged Residues in the Thrombin Structure | p. 15 |
| Thrombin A Chain | p. 17 |
| Thrombin B Chain | p. 19 |
| Thrombin Sites of Specific Function | p. 21 |
| Hirudin | |
| Hirudin-Thrombin Crystal Structure Analysis | p. 38 |
| Nuclear Magnetic Resonance Studies of Thrombin-Fibrinopeptide and Thrombin-Hirudin Complexes | p. 63 |
| NMR Studies | p. 66 |
| NMR Introduction | p. 66 |
| Thrombin-Fibrinopeptide Interaction | p. 67 |
| Thrombin-Hirudin Interaction | p. 75 |
| Preliminary Docking Experiments | p. 79 |
| ESR and Fluorescence Studies of Thrombin Active Site Conformation | p. 87 |
| Biophysical Approaches - ESR Spin Labeling and Fluorescence | p. 88 |
| The Spin Label Technique | p. 88 |
| Fluorescence Spectroscopy | p. 90 |
| Comparing the Active Sites of Human and Bovine Thrombins | p. 92 |
| Human Thrombins | p. 92 |
| Bovine versus Human Thrombin | p. 98 |
| Fluorescent Probes of the Thrombin Active Site | p. 99 |
| Thrombin Complexes with Regulatory Metabolites | p. 103 |
| Apolar Ligands | p. 103 |
| Nucleotide and Pyrophosphate Analogs | p. 105 |
| Probing Conformational Changes in Thrombin-Hirudin Complexes | p. 106 |
| Probing Conformational Changes in Thrombin-Thrombomodulin Complexes | p. 107 |
| Conclusions from Solution versus Crystal Structures | p. 109 |
| Efficacy of Labeling Techniques | p. 109 |
| Precision of Distance Measurements and the Significance of Mapping Binding Regions by Solution Techniques | p. 110 |
| Biochemistry | |
| Synthetic Substrates and Inhibitors of Thrombin | p. 117 |
| Substrates | p. 118 |
| Peptide 4-Nitroanilides | p. 119 |
| Amino Acid and Peptide Thioesters | p. 121 |
| Fluorogenic Substrates | p. 122 |
| Other Types of Substrates | p. 123 |
| Inhibitors | p. 124 |
| Reversible Inhibitors | p. 125 |
| Irreversible Inhibitors | p. 135 |
| Mechanism-Based Inhibitors | p. 139 |
| Specificity of Thrombin Inhibitors | p. 143 |
| Therapeutic Use of Thrombin Inhibitors | p. 143 |
| Regulation of Thrombin by Antithrombin and Heparin Cofactor II | p. 159 |
| Antithrombin | p. 160 |
| Structure and Function | p. 160 |
| Effect of Heparin on the Reaction with Thrombin and Other Clotting Proteinases | p. 168 |
| Physiological Role of Antithrombin and Heparinlike Polysaccharides | p. 188 |
| Modulators of the Reaction of Antithrombin and Heparin with Thrombin and Other Clotting Proteinases | p. 191 |
| Heparin Cofactor II | p. 194 |
| Structure and Function | p. 194 |
| Effect of Glycosaminoglycans on the Reaction with Thrombin | p. 197 |
| Physiological Role | p. 200 |
| Hirudin Interactions with Thrombin | p. 219 |
| Properties of Hirudin | p. 219 |
| Molecular Mechanism of the Interaction of Hirudin with Thrombin | p. 221 |
| Kinetic Mechanism for the Inhibition of Thrombin by Hirudin | p. 223 |
| Interaction Areas on Thrombin and Hirudin | p. 227 |
| Interactions between the N-Terminal Region of Hirudin and the Active-Site Cleft of Thrombin | p. 227 |
| Interactions between the C-Terminal Region of Hirudin, and the Anion-Binding Exosite of Thrombin | p. 232 |
| Structure-Function Relationships in Synthetic Hirudin Peptides | p. 237 |
| Hirudin Peptides as Probes of Thrombin Structure-Function | p. 242 |
| Design of Novel, Hirudin-Based Thrombin Inhibitors | p. 245 |
| Activation of Human Plasma Factor XIII by Thrombin | p. 257 |
| Dual Role of Thrombin in Blood Clotting: Dissecting the Physiological Pathway for Activation of Factor XIII | p. 257 |
| Fibrin Promotes the Proteolytic Activation of Factor XIII by Thrombin, but Cross-Linked Fibrin Blocks this Promoting Effect | p. 259 |
| Factor XIII Can Be Activated without Prior Proteolytic Cleavage | p. 261 |
| Rationalizing the Involvement of Thrombin in the Activation of Factor XIII | p. 262 |
| Physiology | |
| Effect of Chemical Modification of [alpha]-Thrombin on Its Reaction with Platelets and Nucleated Cells | p. 275 |
| Modified Thrombins | p. 277 |
| Effects of [alpha]-Thrombin on Platelets | p. 280 |
| Effects of [alpha]-Thrombin on Nucleated Cells | p. 288 |
| Comparisons of the Effects of [alpha]-Thrombin between Cell Types | |
| [alpha]-Thrombin Binding Components | p. 290 |
| Responses to Nonenzymatically Active Thrombin | p. 290 |
| Receptor Occupancy | p. 291 |
| Modification of Cell Activation by Active-Site-Inhibited [alpha]-Thrombin | p. 291 |
| Preparation of PPACK-Thrombin | p. 292 |
| Effect of PPACK-Thrombin on the Interaction of [alpha]-Thrombin with Platelets | p. 294 |
| Structure-Function Relations of Protein Modifications Distinct from the Catalytic Site | p. 299 |
| Utility of Chemically Modified Thrombins: Thrombosis Intervention and Treatment | p. 300 |
| Unanswered Questions | p. 302 |
| Functional Domains in Thrombin outside the Catalytic Site: Cellular Interactions | p. 315 |
| Thrombin Involvement in Inflammation | p. 315 |
| Mononuclear Phagocyte Chemotaxis | p. 315 |
| Growth-Promoting Activity in Macrophages | p. 318 |
| Thrombin Cell Surface Receptors on Mononuclear Phagocytic Cells | p. 320 |
| Early Events Associated with Mononuclear Phagocyte Activation | p. 321 |
| Thrombin Regulation of Vessel Wall Components | p. 323 |
| Thrombin Immobilization to the Subendothelial ECM | p. 324 |
| Thrombin as a Growth Factor for Smooth Muscle Cells: Nonenzymatic Mode of Action | p. 330 |
| Cell Adhesion to Thrombin | p. 336 |
| Endothelial Cell Adhesion to Thrombin | p. 337 |
| Effect of Synthetic Peptides on EC Attachment to NO[subscript 2]-[alpha]-Thrombin | p. 337 |
| Effect of Specific Thrombin Inhibitors on EC Attachment | p. 339 |
| Effect of NO[subscript 2]-[alpha]-Thrombin on EC Cytoskeletal Organization | p. 339 |
| Attachment of EC to Native [alpha]-Thrombin | p. 342 |
| Concluding Remarks: Functional Domains within Thrombin and Diverse Cellular Interactions | p. 343 |
| Postclotting Cellular Effects of Thrombin Mediated by Interaction with High-Affinity Thrombin Receptors | p. 351 |
| Thrombin as a Growth Factor | p. 351 |
| Stimulation of Cell Proliferation by Trypsin and Thrombin | p. 351 |
| The Search for Specific Thrombin Substrates or Receptors | p. 353 |
| Protease-Nexin Negatively Regulates Thrombin Mitogenesis | p. 354 |
| Characterization of Thrombin Binding Sites | p. 356 |
| Thrombin Domains Involved in Receptor Interaction | p. 356 |
| Molecular Characterization of the Thrombin Receptor | p. 362 |
| Generation of Mitogenic Signals by Receptor Occupancy and Proteolytic Activity | p. 368 |
| Thrombin Effects on Transmembrane Signals in Fibroblasts | p. 368 |
| Mitogenic Signals Generated by Thrombin or DIP-Thrombin Occupancy of Thrombin Receptors | p. 372 |
| Thrombin Interaction with Other Cells: Are the Same Thrombin Domains and Receptors Universally Involved in Cell Activation? | p. 373 |
| Effect of Thrombin on Neuronal Cells | p. 373 |
| Effect of Thrombin on Endothelial Cells | p. 376 |
| Thrombin Effects on Cells of the Inflammatory Response | p. 381 |
| Toward a Strategy for Modulating Wound Healing in Vivo | p. 382 |
| Potential Role of Thrombin in Cellular Aspects of Wound Healing | p. 383 |
| Effects of Thrombin and Thrombin Receptor-Activating Peptides (TRAPs) on Wound Healing in Vivo | p. 385 |
| Regulation of Thrombin-Induced Endothelial Barrier Dysfunction and Prostaglandin Synthesis | p. 397 |
| Thrombin-Mediated Endothelial Cell Barrier Dysfunction | p. 398 |
| Animal Studies of Thrombin-Mediated Lung Edema Formation | p. 399 |
| In Vitro Studies of Thrombin-Induced Endothelial Permeability | p. 403 |
| Thrombin-Stimulated Endothelial Cell Prostaglandin Synthesis | p. 408 |
| Relationship of Thrombin Binding to Prostaglandin Synthesis | p. 408 |
| Thrombin Activation of Arachidonate Releasing Pathways | p. 410 |
| Second Messengers Involved in Thrombin-Mediated Endothelial Cell Effects | p. 410 |
| Regulation of Ca[superscript 2+] in Cultured Endothelial Cells | p. 410 |
| Role in Thrombin-Mediated PGI[subscript 2] Synthesis | p. 411 |
| Role of Ca[superscript 2+] in Thrombin-Mediated Permeability Changes | p. 412 |
| Role Guanine Nucleotide Regulatory Proteins in PGI[subscript 2] Synthesis | p. 416 |
| Role of Protein Kinase C Activation Thrombin-Mediated Prostaglandin Synthesis | p. 420 |
| Role of Protein Kinase C Activation in Thrombin-Mediated Barrier Dysfunction | p. 423 |
| Thrombin's Enzymatic Activity | p. 423 |
| Index | p. 431 |
| Table of Contents provided by Blackwell. All Rights Reserved. |
