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Table of Contents
Preface; Acknowledgments; Contents; 1 Expression Screening of Integral Membrane Proteins by Fusion to Fluorescent Reporters; 1.1 Introduction; 1.2 Bacteria; 1.3 Yeast; 1.4 Insect and Mammalian Cells; 1.5 Summary and Conclusions; References; 2 Detergents in Membrane Protein Purification and Crystallisation; 2.1 Physicochemical Properties of Detergents; 2.2 Role of Detergents in Membrane Protein Solubilisation; 2.3 Detergent Exchange or Removal; 2.4 Role of Detergents in Membrane Protein Stability; 2.5 Detergents for Membrane Protein Purification and Crystallisation
2.5.1 Conventional Detergents2.5.2 New Detergents; 2.5.3 The Role of Detergents in Membrane Protein Crystallisation; 2.5.3.1 Mixed Micelles; 2.6 Conclusions; References; 3 NMR of Membrane Proteins: Beyond Crystals; 3.1 Introduction; 3.2 Barrel Folds; 3.3 Helical Targets; 3.4 Lipid Recognition Domains; 3.5 Protein Structure in Membranes; 3.6 Technologies; 3.7 DNP Sensitivity Enhancement; References; 4 Characterisation of Conformational and Ligand Binding Properties of Membrane Proteins Using Synchrotron Radiation Circular Dichroism (SRCD); 4.1 Introduction
4.2 Diamond B23 Beamline for Synchrotron Radiation Circular Dichroism (SRCD)4.3 Ligand Binding Studies of the FsrC, Ace1 and SbmA Membrane Proteins; 4.4 Protein Stability; 4.5 Temperature Denaturation; 4.6 Conformational Analysis and HT-CD Screening of Protein Crystallisation Buffers; 4.7 Conclusion; References; 5 Membrane Protein Crystallisation: Current Trends and Future Perspectives; 5.1 Introduction; 5.2 Current Trends in the Number and Types of Alpha Helical Membrane Protein Structures; 5.3 Detergent Selection
5.4 Precipitants
How Do They Differ Between Membrane Proteins and Their Soluble Counterparts?5.5 MemGold and MemGold2
What's the Difference?; 5.6 Buffers, pH and Salts; 5.7 MemAdvantage
An Alpha Helical Membrane Protein Additive Screen; 5.8 MemMeso
A Systematically Designed in meso Crystallisation Screen; 5.9 Conclusions; References; 6 Crystal Dehydration in Membrane ProteinCrystallography; 6.1 Introduction; 6.2 The Dehydration Method; 6.2.1 Dehydration by Modifying the Vapour Equilibrium; 6.2.2 Dehydration by Modifying the Chemical Equilibrium
6.2.3 Relative Humidity and Its Relationship with Cryo-Protection6.2.4 Crystal Changes Induced by Dehydration; 6.2.5 Effect of Dehydration on the Membrane-Protein Crystal Lattice; 6.3 Tools and Techniques for Crystal Dehydration; 6.3.1 Dehydration in the Laboratory; 6.3.1.1 Simple Air Exposure; 6.3.1.2 Vapour Control by Exchanging Well Solution; 6.3.1.3 Dehydration by Chemical Exchange of Crystallisation Droplet; 6.3.2 Modern Dehydration Methods with X-ray Feedback; 6.3.2.1 Capillary Dehydration; 6.3.2.2 Dehydration Using the FMS; 6.3.2.3 Dehydration Using the HC1 System
2.5.1 Conventional Detergents2.5.2 New Detergents; 2.5.3 The Role of Detergents in Membrane Protein Crystallisation; 2.5.3.1 Mixed Micelles; 2.6 Conclusions; References; 3 NMR of Membrane Proteins: Beyond Crystals; 3.1 Introduction; 3.2 Barrel Folds; 3.3 Helical Targets; 3.4 Lipid Recognition Domains; 3.5 Protein Structure in Membranes; 3.6 Technologies; 3.7 DNP Sensitivity Enhancement; References; 4 Characterisation of Conformational and Ligand Binding Properties of Membrane Proteins Using Synchrotron Radiation Circular Dichroism (SRCD); 4.1 Introduction
4.2 Diamond B23 Beamline for Synchrotron Radiation Circular Dichroism (SRCD)4.3 Ligand Binding Studies of the FsrC, Ace1 and SbmA Membrane Proteins; 4.4 Protein Stability; 4.5 Temperature Denaturation; 4.6 Conformational Analysis and HT-CD Screening of Protein Crystallisation Buffers; 4.7 Conclusion; References; 5 Membrane Protein Crystallisation: Current Trends and Future Perspectives; 5.1 Introduction; 5.2 Current Trends in the Number and Types of Alpha Helical Membrane Protein Structures; 5.3 Detergent Selection
5.4 Precipitants
How Do They Differ Between Membrane Proteins and Their Soluble Counterparts?5.5 MemGold and MemGold2
What's the Difference?; 5.6 Buffers, pH and Salts; 5.7 MemAdvantage
An Alpha Helical Membrane Protein Additive Screen; 5.8 MemMeso
A Systematically Designed in meso Crystallisation Screen; 5.9 Conclusions; References; 6 Crystal Dehydration in Membrane ProteinCrystallography; 6.1 Introduction; 6.2 The Dehydration Method; 6.2.1 Dehydration by Modifying the Vapour Equilibrium; 6.2.2 Dehydration by Modifying the Chemical Equilibrium
6.2.3 Relative Humidity and Its Relationship with Cryo-Protection6.2.4 Crystal Changes Induced by Dehydration; 6.2.5 Effect of Dehydration on the Membrane-Protein Crystal Lattice; 6.3 Tools and Techniques for Crystal Dehydration; 6.3.1 Dehydration in the Laboratory; 6.3.1.1 Simple Air Exposure; 6.3.1.2 Vapour Control by Exchanging Well Solution; 6.3.1.3 Dehydration by Chemical Exchange of Crystallisation Droplet; 6.3.2 Modern Dehydration Methods with X-ray Feedback; 6.3.2.1 Capillary Dehydration; 6.3.2.2 Dehydration Using the FMS; 6.3.2.3 Dehydration Using the HC1 System