Handbook of Synthetic Substrates

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.- IV.2. Factor Xa and derived determinations.- IV.3. Determination of fibrinolytic pathway components in plasma.- IV.4. The contact activation system.- IV.5. Miscellaneous determinations.- Bibliography of synthetic substrates (as available Fall 1982).- Index of subjects.