Handbook of Synthetic Substrates

H.C. Hemker (Autor)

Buch | Hardcover

192 Seiten

1983 | 1983 ed.
Kluwer Academic Publishers (Verlag)
978-0-89838-556-4 (ISBN)

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The need for a handbook on the use of synthetic substrates for assay of proteases of the coagulation and fibrinolytic systems became evident several years ago during the activities of the Subcommittee on Synthetic Substrates of the International Committee on Thrombosis and Haemo- stasis (lCTH). Production of such a handbook, which was recommended during discussions of the ICTH at its meeting in London in 1979 was made possible by the generous efforts of Professor HC Hemker with the aid of several contributors with particular interests in the use of synthetic substrates in coagulation and fibrinolysis. As current Chairman and Secretary General of the ICTH we would like to express our sincere thanks to Professor Hemker for producing this handbook and look for- ward to seeing the benefits of this tremendous effort reflected in the advancement of our understanding of thrombosis and hemostasis and the transfer of such knowledge into improved diagnosis and treatment of thrombotic and hemorrhagic disorders. Craig M Jackson Professor of Biological Chemistry Washington University School of Medicine, St. Louis, MO Chairman, ICTH Harold R Roberts Professor of Internal Medicine University of North Carolina, Chapel Hill, NC Secretary General, ICTH ix Foreword The advent of synthetic substrates for the study of blood coagulation and fibrinolysis was a significant step forward in the investigation of these systems. Both basic research and clinical laboratory investigations can profit from these advanced tools.

I. Basic enzymology.- I. 1. Introduction.- I. 2. Maximal velocity.- I. 3. Michaelis constant.- I. 4. Low substrate concentration; the efficiency of an enzyme.- I. 5. High substrate concentration; the saturation curve.- I. 6. The characteristic parameters of an enzymatic reaction.- I. 7. Enzyme inhibition.- I. 8. Rate-constants and enzymatic mechanism.- I. 9. The reaction mechanism of serine proteases.- 1.10. Transient state kinetics.- 1.11. Data from a reaction going to completion.- 1.12. Active site titration.- 1.13. Simultaneous reactions in complicated reaction mixtures.- II. Measuring the conversion of a chromogenic substrate.- II. 1. Photometric assays.- II. 2. Absorption spectrophotometry.- II. 3. Choice of wavelength and absorbancy range.- II. 4. Temperature.- II. 5. The reaction medium.- II. 6. Substrate solution.- II. 7. Methodology.- II. 8. Execution of the experiment.- II. 9. Fluorescence.- II.10. Electrochemical determinations.- III. Substrates.- III.1. Specificity in serine proteases.- III.2. Substrate selectivity and sensitivity.- III.3. Inhibition by artificial substrates.- III.4. Data on commercially available substrates.- III.5. Fluorogenic substrates.- III.6. General principles of fluorogenic peptide substrates with 7-amino-4-methylcoumarine (MCA) as a leaving group.- III.7. Thioesters.- III.8. Substrates for electrochemical determinations.- III.9. Luminogenic substrates.- IV. Determinations that can be carried out with chromogenic substrates.- IV. 1. Thrombin and derived determinations.- 1.1. thrombin.- 1.2. prothrombin.- 1.3. antithrombin III and heparin cofactor.- 1.4. prothrombinase, factor V, platelet factor 3.- 1.4.1. prothrombinase.- 1.4.2. factor V.- 1.4.3. platelet factor 3.- IV.2. Factor Xa and derived determinations.- 2.1. factor Xa.- 2.2. factor X.- 2.3. antifactor Xa (antithrombin III, heparin cofactor).- 2.4. heparin.- 2.5. platelet factor 4 and other antiheparins.- 2.6. factor X activating enzymes.- 2.6.1. factor VIII.- 2.6.2. factor IX.- 2.6.3. platelet factor 3.- 2.6.4. factor VII and tissue thromboplastin.- IV.3. Determination of fibrinolytic pathway components in plasma.- 3.1. plasminogen.- 3.2. plasminogen activator.- 3.3. `free’ protease activity.- 3.4. plasmin generation rates.- 3.5. ?2-plasmin inhibitor.- IV.4. The contact activation system.- 4.1. plasma prekallikrein.- 4.2. factor XII.- 4.3. activated factor XI.- IV.5. Miscellaneous determinations.- 5.1. glandular kallikrein.- 5.2. brinase and brinase inhibitors.- 5.3. bacterial endotoxins.- Bibliography of synthetic substrates (as available Fall 1982).- Index of subjects.

Erscheint lt. Verlag	30.6.1983
Zusatzinfo	192 p.
Verlagsort	Dordrecht
Sprache	englisch
Gewicht	450 g
Themenwelt	Medizinische Fachgebiete ► Innere Medizin ► Hämatologie
ISBN-10	0-89838-556-3 / 0898385563
ISBN-13	978-0-89838-556-4 / 9780898385564
Zustand	Neuware