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Measurement and analysis of kinetic isotope effects /
This volume serves as a guide to new developments that take advantage of the increased instrumental capabilities that have become available in recent years. It also reflects the expansion of kinetic isotope effects into the environment and drug metabolism fields. Newer methods from higher resolution...
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | , |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Cambridge, MA :
Academic Press is an imprint of Elsevier,
2017.
|
| Colección: | Methods in enzymology ;
v. 596. |
| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- Front Cover; Measurement and Analysis of Kinetic Isotope Effects; Copyright; Contents; Contributors; Preface; References; Chapter One: Binding Isotope Effects for Interrogating Enzyme-Substrate Interactions; 1. Introduction; 2. Equipment; 3. Materials; 3.1. Buffers; 3.2. Radiolabeled Substrates; 4. Measurement of Equilibrium BIEs Using the RED Device; 4.1. Determining Substrate Equilibration Time; 4.2. Determination of Substrate Stability; 4.3. Measurement of BIEs on Binary Complex Association; 4.4. Measurement of BIEs on Ternary Complex Association
- 4.5. Scintillation Counting and Spectral Deconvolution5. BIE Interpretation and Examples; 6. Conclusions; Acknowledgments; References; Chapter Two: Chemical Ligation and Isotope Labeling to Locate Dynamic Effects; 1. Introduction; 2. A General Overview of the Approach; 2.1. Production of N-Terminal Thioester Peptide EcDHFR (1-28); 2.2. Production of C-Terminal Cysteine Peptide EcDHFR-A29C (29-159); 2.3. Chemical Ligation to Form Full-Length EcDHFR; 2.4. Peptide Mass Spectrometry; 2.5. Circular Dichroism Spectroscopy; 2.6. Steady-State Kinetic Measurements
- 2.3. Effect of Enzyme Isotopic Labeling on Catalysis2.3.1. Effect on Steady-State Kinetic Parameters; 2.3.2. Effect on Chemical Step; 2.3.2.1. Computational Studies; 2.3.2.2. Experimental Studies: Enzyme Isotope Effects on the Rate of the Chemical Step; 2.3.2.3. Experimental Studies: Enzyme Isotope Effects on Substrate Intrinsic Kinetic Isotope Effects; 2.3.3. Regio-Specific Effects of Enzyme Isotopic Labeling on Catalysis; 3. Summary; Acknowledgments; References; Chapter Four: Extracting Kinetic Isotope Effects From a Global Analysis of Reaction Progress Curves; 1. Introduction
- 2. (Global) Nonlinear Fitting3. Global Fitting of a Time Course Described by Integrated Rate Equations; 4. Modeling and Fitting Time Courses Using Numerical Integration of Rate Equations; 5. Model Selection and Error Analysis; 6. Specific Examples; 6.1. DNA Polymerase; 6.2. Ethanolamine Ammonia Lyase; 6.3. Model-Free Calculation of KIEs; 7. Summary; Acknowledgments; References; Chapter Five: Natural Isotope Abundance in Metabolites: Techniques and Kinetic Isotope Effect Measurement in Plant, Anima ... ; 1. Introduction; 2. Compound-Specific 13C and 15N Analysis: History and Overview