Enzymology

  1. Enzyme Kinetics
    1. Fundamentals of Chemical Kinetics
      1. Reaction Rates and Rate Laws
        1. Definition of Rate
          1. Rate Constants
            1. Factors Affecting Reaction Rates
            2. Reaction Order
              1. Zero-Order Reactions
                1. First-Order Reactions
                  1. Second-Order Reactions
                    1. Pseudo-Order Reactions
                  2. Measuring Enzyme Activity
                    1. Enzyme Assays
                      1. Continuous Assays
                        1. Discontinuous Assays
                          1. Spectrophotometric Assays
                            1. Fluorometric Assays
                              1. Radiometric Assays
                                1. Coupled Assays
                                2. Definition of Initial Velocity
                                  1. Importance in Kinetic Analysis
                                    1. Linear Phase of Reaction
                                    2. Units of Enzyme Activity
                                      1. International Unit
                                        1. Katal
                                          1. Specific Activity
                                            1. Molar Activity
                                          2. Single-Substrate Enzyme Kinetics
                                            1. The Michaelis-Menten Equation
                                              1. Derivation and Assumptions
                                                1. Steady-State Assumption
                                                  1. Rapid Equilibrium Assumption
                                                    1. Pre-Equilibrium Approximation
                                                    2. Michaelis Constant
                                                      1. Interpretation and Significance
                                                        1. Relationship to Binding Affinity
                                                        2. Maximum Velocity
                                                          1. Determination and Meaning
                                                            1. Relationship to Enzyme Concentration
                                                          2. Catalytic Efficiency
                                                            1. kcat/Km Ratio
                                                              1. Diffusion Limit
                                                                1. Enzyme Perfection
                                                                2. Turnover Number
                                                                  1. Definition and Calculation
                                                                    1. Biological Significance
                                                                  2. Graphical Analysis of Kinetic Data
                                                                    1. Michaelis-Menten Plot
                                                                      1. Hyperbolic Curve
                                                                        1. Parameter Estimation
                                                                        2. Lineweaver-Burk Plot
                                                                          1. Double Reciprocal Plot
                                                                            1. Linearization of Data
                                                                              1. Limitations and Artifacts
                                                                              2. Eadie-Hofstee Plot
                                                                                1. Advantages and Disadvantages
                                                                                  1. Error Distribution
                                                                                  2. Hanes-Woolf Plot
                                                                                    1. Interpretation
                                                                                      1. Statistical Considerations
                                                                                      2. Non-Linear Regression Analysis
                                                                                      3. Enzyme Inhibition
                                                                                        1. Reversible Inhibition
                                                                                          1. Competitive Inhibition
                                                                                            1. Mechanism and Kinetic Effects
                                                                                              1. Overcoming by Substrate
                                                                                                1. Inhibition Constant
                                                                                                2. Uncompetitive Inhibition
                                                                                                  1. Mechanism and Kinetic Effects
                                                                                                    1. Kinetic Parameters
                                                                                                    2. Non-competitive Inhibition
                                                                                                      1. Mechanism and Kinetic Effects
                                                                                                        1. Pure vs Mixed Non-competitive
                                                                                                        2. Mixed Inhibition
                                                                                                          1. Mechanism and Kinetic Effects
                                                                                                            1. Alpha and Beta Factors
                                                                                                          2. Irreversible Inhibition
                                                                                                            1. Covalent Modification of Active Site
                                                                                                              1. Time-Dependent Inhibition
                                                                                                                1. Examples of Irreversible Inhibitors
                                                                                                                  1. Suicide Inhibitors
                                                                                                                    1. Mechanism-Based Inactivators
                                                                                                                      1. Mechanism of Action
                                                                                                                  2. Kinetics of Multi-Substrate Reactions
                                                                                                                    1. Sequential Mechanisms
                                                                                                                      1. Ordered Sequential
                                                                                                                        1. Substrate Binding Order
                                                                                                                          1. Kinetic Patterns
                                                                                                                          2. Random Sequential
                                                                                                                            1. Flexible Binding Order
                                                                                                                              1. Dead-End Complexes
                                                                                                                            2. Ping-Pong Mechanism
                                                                                                                              1. Double-Displacement Mechanism
                                                                                                                                1. Enzyme Intermediate Formation
                                                                                                                                  1. Kinetic Characteristics
                                                                                                                                  2. Product Inhibition Studies
                                                                                                                                    1. Isotope Exchange Studies

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                                                                                                                                  3. Principles of Enzyme Catalysis

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                                                                                                                                  5. Regulation of Enzyme Activity