Isothermal Titration Calorimetry in Enzymology : Techniques and Applications.

Format:

Book

Author/Creator:

Mittermaier, Anthony.

Contributor:

Di Trani, Justin.

Series:

Foundations and Frontiers in Enzymology Series

Language:

English

Physical Description:

1 online resource (532 pages)

Edition:

1st ed.

Place of Publication:

Chantilly : Elsevier Science & Technology, 2025.

Summary:

Isothermal Titration Calorimetry in Enzymology: Techniques and Applications provides a thorough, practical overview of ITC as a productive and powerful tool for quantifying enzyme catalysis and interactions with substrates, products, cofactors, and inhibitors.

Contents:

Front Cover

Isothermal Titration Calorimetry in Enzymology

Isothermal Titration Calorimetry in Enzymology: Techniques and Applications

Copyright

Contents

Contributors

Preface

I-ITC kinetics measurements

1 - kinITC: When thermodynamics meets kinetics

1. Introduction

2. The instrument response time(s)

3. Experimental example of signal deconvolution with one response time

4. The different methods to obtain kinetic information

5. Quality of the response time obtained by fitting the ETC

6. Theoretical and practical considerations about injection curve fitting

7. Comparison of full kinITC with one and two response times

Appendix

References

2 - Characterizing enzyme kinetics by 2D-ITC

2. Instrumentation and the empirical response model (ERM)

3. Measuring enzymes kinetics with ITC

4. Performing a multiple injection experiment

5. Performing injections of substrate and inhibitor into enzyme

6. Enzymes with two substrates

7. The enzyme velocity surface

8. Using 2D ITC to sample the enzyme velocity surface

9. Model determination and parameter fitting with 2D-ITC

10. Conclusions

11. Appendix

3 - Using isothermal titration calorimetry to measure enzyme stability

1. Enzymes in industry

2. Enzyme structure and stability

3. Current methods for measuring protein stability

4. Measuring enzyme stability with isothermal titration calorimetry (ITC)

5. Conclusions

4 - Analysis of ITC enzyme kinetics data using the Lambert W function

2. The calorimetric signal

3. The integrated Michaelis-Menten equation

4. Experimental data analysis

5. Competitive inhibition

6. Computer simulations

7. Experimental design

8. Conclusions

Further reading.

5 - Design and operation of small-volume isothermal titration calorimeters (ITCs)

2. Designing an ITC

3. Heat measurement

4. Titration

5. Calibration

6. Operation

7. Data analysis

8. Common pitfalls

9. Conclusions

II-ITC thermodynamic

6 - The binding polynomial: A powerful tool for modeling, analyzing, and interpreting isothermal titration calorime ...

2. The partition function: A not-so-distant concept

3. The binding polynomial

4. Properties of the binding polynomial

5. The binding equations: Chemical equilibrium and mass conservation

6. Isothermal titration calorimetry vs. other binding techniques

7. The simplest case: One ligand binding site

8. A more complex case: Two binding sites

9. Other cases (I): Conformational heterogeneity

10. Other cases (II): Heterotropic interactions

11. Conclusions

Acknowledgments

Funding

7 - Unbiased baseline determination, global analysis, and multimethod analysis of titration isotherms in SEDPHAT an ...

2. Thermogram analysis in NITPIC

2.1 Basic problem and strategy

2.2 The NITPIC algorithm

2.3 Practical application of NITPIC and extensions

3. Titration binding isotherm analysis in SEDPHAT

3.1 Basic organization

3.2 Simultaneous analysis of multiple ITC titration isotherms

3.3 Multisite interactions

3.4 Global multimethod analysis

4. Discussion

8 - Characterization of slow-binding inhibition by isothermal titration calorimetry: The case of urease, a nickel-d ...

1. An overview on enzyme slow-binding inhibition

2. Slow-binding inhibition kinetics determined by ITC

2.1 ITC overview and instrumentation

2.2 Enzyme kinetics using ITC

3. Characterization of urease slow-binding inhibition using ITC.

4. Summary

9 - Dissecting complex binding equilibria involving protein-protein and protein-metal interactions

2. Theoretical background

3. Exploring protein-protein and protein-metal interactions for Ni(II) delivery into urease

4. Insights into the role of calmodulin as a Ca(II)-dependent adaptor protein

5. Study of a metal-induced conformational change in Ni(II)-superoxide dismutase

6. Conclusions

10 - Thermodynamics of transition state analogs

1. Enzymatic transition state structures and transition state analogs

2. Isotope effects and enzymatic transition states

3. Thermodynamic signatures of transition state analogs

4. Transition states of N-ribosyltransferases N-ribosylhydrolases and associated transition state analogs

5. Purine nucleoside phosphorylase

5.1 The importance of PNP as a biological target

5.2 Transition state analysis of PNP

5.3 Inhibition of PNP with transition state analogs

5.4 Thermodynamics of immucillins binding to PNP

6. 5ʹ-Methylthioadenosine phosphorylase

6.1 The importance of MTAP as a biological target

6.2 Transition state structure of human MTAP and transition state analogs

6.3 Thermodynamics of MTAP transition state analog binding

7. 5ʹ-Methylthioadenosine nucleosidases

7.1 The importance of MTAN's as antibiotic targets

7.2 MTAN transition states and inhibition of MTANs by transition state analogs

7.3 Thermodynamics of MTAN transition state analog binding

7.4 Structural and molecular dynamics analysis of transition state analog binding to MTANs

8. Summary

11 - Application of isothermal titration calorimetry in drug discovery and development

2. Application of ITC in initial screening of binders.

2.1 Application of ITC to evaluate ligand binding ability of proteins

2.2 Application of ITC for fragment screening

3. Application of ITC in hit validation and characterization

3.1 Application of ITC in determining accurate binding affinity

3.2 Utilization of thermodynamic binding data given by ITC

3.3 Measurement of kinetic data using ITC

4. Application of ITC in hit-to-lead-to-drug optimization

12 - Insight on metalloenzymes from isothermal titration calorimetry measurements

2. Hydrolases

2.1 Nucleases

2.2 Aminopeptidases

2.3 Phosphatases

2.4 Metallo-β-lactamase

2.5 Carbonic anhydrase

2.6 Ribozymes

3. Dioxygenases

3.1 Taurine dioxygenase

3.2 Ethylene-forming enzyme

3.3 Aspartyl(asparaginyl)-β-hydroxylase

4. Other metalloenzymes

4.1 Homoprotocatechuate-2,3-dioxygenase

4.2 Tartrate dehydrogenase

13 - Thermophilicity, substrate promiscuity, and solvent effects on thermodynamics of ligand-protein interactions s ...

2. Ligand binding to enzymes with different levels of substrate promiscuity

3. Ternary complexes

4. Determination of solvent effects in ligand binding

5. Conclusions and future prospects

14 - Isothermal titration calorimetry: A thermodynamic window into natural product binding with cancer targets

2. Anticancer potential of natural products

3. Overexpression of regulatory proteins in cancer

4. ITC in cancer therapeutics

4.1 The advantage of ITC in cancer drug development

4.2 Measurement of thermodynamic parameters

5. Interpretation of ITC in terms of ligand-protein interaction

15 - ITC of intrinsically disordered proteins

1. Introduction.

2. Intrinsically disordered proteins

3. ITC of IDPs

3.1 Probing coupled conformational transitions in interacting IDPs with ITC

3.2 Evaluating conformational entropy in IDR-based molecular recognition

3.3 Illustrative examples of using ITC for the IDP analysis

Index

Back Cover.

Notes:

Description based on publisher supplied metadata and other sources.

ISBN:

0-443-21849-8

9780443218491

OCLC:

1545645538