Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
This third edition provides new and updated chapters detailing a complete introduction to common and emerging procedures for characterizing the interactions of individual proteins with their natural ligands, drugs or other binding partners. Chapters detail natural substrates, potential drug leads, quantitative understanding of the mechanism of interaction, and different techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Protein Ligand Interactions: Methods and Applications, Third Edition serves as an ideal guide for researchers new to the field of biophysical characterization of protein interactions.
Part I : Overview
1. Assessing and Improving Protein Sample Quality
Bertrand Raynal, Sébastien Brûlé, Stephan Uebel, and Stefan H. Knauer
2. A Familiar Protein-Ligand Interaction Revisited with Multiple Methods
Xiaochun Li-Blatter, Ludovit Zweifel, and Timothy Sharpe
Part II: Universal Methods For Protein Interactions
3. Interactions of a Signal Transduction Protein Investigated by Fluorescence Stopped-Flow Kinetics
Stephen R. Martin and Maria J. Schilstra
4. Kinetic Methods of Deducing Binding Mechanisms Involving Intrinsically Disordered Proteins
Elin Karlsson and Per Jemth
5. Isothermal Titration Calorimetry
Christopher M. Johnson
6. Measuring the K
d of Protein-Ligand Interactions Using Microscale Thermophoresis
Shih-Chia Tso and Chad A. Brautigam
7. Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D); Preparing Functionalized Lipid Layers for the Study of Complex Protein-Ligand Interactions
Holly L Birchenough and Thomas A Jowitt
Part III: Screening For Ligand Binding
8. Indirect Detection of Ligand Binding by Thermal Melt Analysis Joseph Shaw and Christopher Stubbs
9. The Use of Acoustic Mist Ionization Mass Spectrometry for High-throughput Screening
Helen Plant, David Murray, Hannah Semple, Gareth Davies, Ian Sinclair, and Geoffrey A. Holdgate
10. Ligand Discovery – High Through-put Binding: Fluorescence Polarization (Anisotropy)
Geoffrey A. Holdgate and Paul E. Hemsley
11. Fragment Screening by NMR
Ben J. Davis
Part IV: Nucleotide Binding and Hydrolysis
12. A Quick Primer in Fluorescence-based Equilibrium and Pre-Steady State Methods for Determining Protein-Nucleotide Affinities
Harland E. Brandon and Hans-Joachim Wieden
13. Measurement of Nucleotide Hydrolysis using Fluorescent Biosensors for Phosphate
Simone Kunzelmann
Part V: Binding Nucleic Acids
14. Gel-based Analysis of Protein-Nucleic Acid Interactions
James A.W. Stowell, Terence T.L. Tang, Maximilian Seidel, and Lori A. Passmore
15. Biophysical Studies of the Binding of Viral RNA with the 80S Ribosome using SwitchSENSE
Emma Schenckbecher, Guillaume Bec, Taiichi Sakamoto, Benoit Meyer, and Eric Ennifar
16. Biolayer Interferometry: Protein-RNA Interactions
Stephen R. Martin, Andres Ramos, and Laura Masino
17. Analysis of Protein-DNA Interactions Using Surface Plasmon Resonance and a ReDCaT Chip
Clare E. M. Stevenson and David M. Lawson
18. Characterization of Protein Nucleic Acid complexes by Size-Exclusion Chromatography Coupled with Light Scattering, Absorbance and Refractive Index Detectors
Ewa Folta-Stogniew
19. Analytical Ultracentrifugation for Analysis of Protein Nucleic Acid Interactions
Andrea Bogutzki and Ute Curth
20. Studying RNA-Protein Complexes Using X-ray Crystallography
Andrew P. Turnbull and Xiaoqiu Wu
Part VI: Membrane Binding
21. Flow Linear Dichroism of Protein-membrane Systems
Matthew R.Hicks, Sarah R. Dennison, Adewale Olamoyesan, and Alison Rodger
22. Probing Protein-Membrane Interactions and Dynamics Using Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS)
Jordan TB Stariha, Reece M Hoffmann, David J Hamelin, and John E Burke
This third edition provides new and updated chapters detailing a complete introduction to common and emerging procedures for characterizing the interactions of individual proteins with their natural ligands, drugs or other binding partners. Chapters detail natural substrates, potential drug leads, quantitative understanding of the mechanism of interaction, and different techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Protein Ligand Interactions: Methods and Applications, Third Edition serves as an ideal guide for researchers new to the field of biophysical characterization of protein interactions.
Assessing and Improving Protein Sample Quality.- A Familiar Protein-Ligand Interaction Revisited with Multiple Methods.- Interactions of a Signal Transduction Protein Investigated by Fluorescence Stopped-Flow Kinetics.- Kinetic Methods of Deducing Binding Mechanisms Involving Intrinsically Disordered Proteins.- Isothermal Titration Calorimetry.- Measuring the Kd of Protein-Ligand Interactions Using Microscale Thermophoresis.- Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D); Preparing Functionalized Lipid Layers for the Study of Complex Protein-Ligand Interactions.- Indirect Detection of Ligand Binding by Thermal Melt Analysis.- The Use of Acoustic Mist Ionization Mass Spectrometry for High-throughput Screening.- Ligand Discovery - High Through-put Binding: Fluorescence Polarization (Anisotropy).- Fragment Screening by NMR.- A Quick Primer in Fluorescence-based Equilibrium and Pre-Steady State Methods for Determining Protein-Nucleotide Affinities.- Measurement of Nucleotide Hydrolysis using Fluorescent Biosensors for Phosphate.- Gel-based Analysis of Protein-Nucleic Acid Interactions.- Biophysical Studies of the Binding of Viral RNA with the 80S Ribosome using SwitchSENSE.- Biolayer Interferometry: Protein-RNA Interactions.- Analysis of Protein-DNA Interactions Using Surface Plasmon Resonance and a ReDCaT Chip.- Characterization of Protein Nucleic Acid complexes by Size-Exclusion Chromatography Coupled with Light Scattering, Absorbance and Refractive Index Detectors.- Analytical Ultracentrifugation for Analysis of Protein Nucleic Acid Interactions.- Studying RNA-Protein Complexes Using X-ray Crystallography.- Flow Linear Dichroism of Protein-membrane Systems.- Probing Protein-Membrane Interactions and Dynamics Using Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS).
Inhaltsverzeichnis
Part I : Overview
1. Assessing and Improving Protein Sample Quality
Bertrand Raynal, Sébastien Brûlé, Stephan Uebel, and Stefan H. Knauer
2. A Familiar Protein-Ligand Interaction Revisited with Multiple Methods
Xiaochun Li-Blatter, Ludovit Zweifel, and Timothy Sharpe
Part II: Universal Methods For Protein Interactions
3. Interactions of a Signal Transduction Protein Investigated by Fluorescence Stopped-Flow Kinetics
Stephen R. Martin and Maria J. Schilstra
4. Kinetic Methods of Deducing Binding Mechanisms Involving Intrinsically Disordered Proteins
Elin Karlsson and Per Jemth
5. Isothermal Titration Calorimetry
Christopher M. Johnson
6. Measuring the K
d of Protein-Ligand Interactions Using Microscale Thermophoresis
Shih-Chia Tso and Chad A. Brautigam
7. Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D); Preparing Functionalized Lipid Layers for the Study of Complex Protein-Ligand Interactions
Holly L Birchenough and Thomas A Jowitt
Part III: Screening For Ligand Binding
8. Indirect Detection of Ligand Binding by Thermal Melt Analysis Joseph Shaw and Christopher Stubbs
9. The Use of Acoustic Mist Ionization Mass Spectrometry for High-throughput Screening
Helen Plant, David Murray, Hannah Semple, Gareth Davies, Ian Sinclair, and Geoffrey A. Holdgate
10. Ligand Discovery - High Through-put Binding: Fluorescence Polarization (Anisotropy)
Geoffrey A. Holdgate and Paul E. Hemsley
11. Fragment Screening by NMR
Ben J. Davis
Part IV: Nucleotide Binding and Hydrolysis
12. A Quick Primer in Fluorescence-based Equilibrium and Pre-Steady State Methods for Determining Protein-Nucleotide Affinities
Harland E. Brandon and Hans-Joachim Wieden
13. Measurement of Nucleotide Hydrolysis using Fluorescent Biosensors for Phosphate
Simone Kunzelmann
Part V: Binding Nucleic Acids
14. Gel-based Analysis of Protein-Nucleic Acid Interactions
James A.W. Stowell, Terence T.L. Tang, Maximilian Seidel, and Lori A. Passmore
15. Biophysical Studies of the Binding of Viral RNA with the 80S Ribosome using SwitchSENSE
Emma Schenckbecher, Guillaume Bec, Taiichi Sakamoto, Benoit Meyer, and Eric Ennifar
16. Biolayer Interferometry: Protein-RNA Interactions
Stephen R. Martin, Andres Ramos, and Laura Masino
17. Analysis of Protein-DNA Interactions Using Surface Plasmon Resonance and a ReDCaT Chip
Clare E. M. Stevenson and David M. Lawson
18. Characterization of Protein Nucleic Acid complexes by Size-Exclusion Chromatography Coupled with Light Scattering, Absorbance and Refractive Index Detectors
Ewa Folta-Stogniew
19. Analytical Ultracentrifugation for Analysis of Protein Nucleic Acid Interactions
Andrea Bogutzki and Ute Curth
20. Studying RNA-Protein Complexes Using X-ray Crystallography
Andrew P. Turnbull and Xiaoqiu Wu
Part VI: Membrane Binding
21. Flow Linear Dichroism of Protein-membrane Systems
Matthew R.Hicks, Sarah R. Dennison, Adewale Olamoyesan, and Alison Rodger
22. Probing Protein-Membrane Interactions and Dynamics Using Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS)
Jordan TB Stariha, Reece M Hoffmann, David J Hamelin, and John E Burke
Klappentext
This third edition provides new and updated chapters detailing a complete introduction to common and emerging procedures for characterizing the interactions of individual proteins with their natural ligands, drugs or other binding partners. Chapters detail natural substrates, potential drug leads, quantitative understanding of the mechanism of interaction, and different techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Protein Ligand Interactions: Methods and Applications, Third Edition serves as an ideal guide for researchers new to the field of biophysical characterization of protein interactions.
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
