Kooperativer Bibliotheksverbund

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Umfang: 1 online resource (531 p.)

ISBN: 0-12-802907-2

Serie: Methods in Enzymology, Volume 567

Originaltitel: Calorimetry (Feig)

Anmerkung: Description based upon print version of record. , Front Cover; Calorimetry; Copyright; Contents; Contributors; Preface; References; Section I: ITC; Chapter One: Isothermal Titration Calorimetry Measurements of Metal Ions Binding to Proteins; 1. Introduction; 2. Metal Ion Properties Relevant to ITC Measurements; 2.1. Aqueous Solution Chemistry of Metal Ions; 2.2. Buffers; 2.3. Protons; 3. ITC Measurements Involving Metal Ions; 4. Fitting ITC Data Involving Metal Ions; 5. Post Hoc Analysis of ITC Data Involving Metal Ions; 5.1. Binding Enthalpy (ΔH); 5.2. Equilibrium Constant (K); 5.3. Binding Entropy (ΔS) and Heat Capacity (ΔCP) , 6. Additional Considerations with Metal Ions6.1. Comparison with Other Methods; 6.2. Sources of Error; 6.3. Tips; 7. Conclusion; Acknowledgments; References; Chapter Two: Assessing Coupled Protein Folding and Binding Through Temperature-Dependent Isothermal Titration Calorimetry; 1. Introduction; 2. Sample Preparation; 2.1. Plasmid Construction; 2.2. Purification of FCP1 and Rap74; 3. Special Considerations for ITC of Intrinsically Disordered Proteins; 3.1. Experimental Design; 3.2. Calorimetric Parameters; 4. Data Fitting and Analysis; 4.1. Standard Data Fitting in Origin , 4.2. Data Fitting in MATLAB4.3. Global Analysis of Temperature-Dependent Data Sets; 5. Summary; Acknowledgments; References; Chapter Three: Fragment-Based Screening for Enzyme Inhibitors Using Calorimetry; 1. Introduction; 1.1. Nanocalorimeter Arrays; 1.2. Fragment-Based Lead Discovery; 2. Required Materials; 2.1. Equipment; 2.2. Reagents; 2.3. Software; 3. Instrumentation; 4. Fragment Library Selection; 4.1. Materials Needed; 4.2. Software Needed; 4.3. Protocol to Prepare of Fragment Library for Screening; 5. Calorimetric Enzyme Assay: Nanocalorimeter , 5.1. Sample Considerations and Preparation5.2. Data Analysis; 5.3. Protocol; 6. Calorimetric Enzyme Assay: Commercial Microcalorimeters; 6.1. Required Materials; 6.2. Cleaning Before and After Runs; 6.3. Protocol 2: Microcalorimeter; 6.4. Data Processing and Analysis; 7. Summary; Acknowledgments; References; Chapter Four: Measuring Multivalent Binding Interactions by Isothermal Titration Calorimetry; 1. Introduction; 2. Determining the Thermodynamics of Lectin Binding to Multivalent Synthetic Analogs; 2.1. Methods; 2.1.1. Isothermal Titration Calorimetry , 2.1.2. Determination of Functional Valence of Multivalent Ligands2.2. Data Analysis and Interpretation; 2.2.1. ITC can Reliably Measure the Enhanced Affinities of Multivalent Carbohydrates; 2.2.2. Information Obtained from ITC-Driven n Values; 2.2.3. Structural Valence of a Multivalent Ligand May Differ from Its Functional Valence; 2.2.4. Macroscopic-Binding Enthalpy of Multivalent Carbohydrate Analogs Is the sum of the Microscopic Enthalpy of the Epi ...; 2.2.5. Macroscopic TΔS of High-Affinity Carbohydrate Analogs Is More Than the Sum of Microscopic TΔS , 2.2.6. Favorable TΔS Contributes to the Enhanced Affinities of Multivalent Carbohydrates

Weitere Ausg.: ISBN 0-12-802906-4

Sprache: Englisch