This second edition of Membrane Protein Purification and Crystallization, A Practical Guide is written for bench scientists working in the fields of biochemistry, biology, and proteomic research. This guide presents isolation and crystallization techniques in a concise form, emphasizing the critical aspects unique to membrane proteins. It explains the principles of the methods and provides protocols of general use, permitting researchers and students new to this area to adapt these techniques to their particular needs. This edition is not only an update but is comprised mainly of new contributions. It is the first monograph compiling the essential approaches for membrane protein crystallization, and emphasizes recent progress in production and purification of recombinant membrane proteins.
Key features
* Provides general guidelines and strategies for isolation and crystallization of membrane proteins
* Gives detailed protocols that have wide application, and low specialized equipment needs
* Emphasizes recent progress in production and purification of recombinant membrane proteins, especially of histidine-tagged and other affinity-epitope-tagged proteins
* Summarizes recent developments of Blue-Native PAGE, a high resolution separation technique, which is independent of the use of recombinant techniques, and is especially suited for proteomic analyses of membrane protein complexes
* Gives detailed protocols for membrane protein crystallization, and describes the production and use of antibody fragments for high resolution crystallization
* Presents a comprehensive guide to 2D-crystallization of membrane proteins
1. Cover image - p. Cover 2. Title page - p. Cover 3. Table of Contents - p. Cover 4. Copyright - p. Cover 5. Contributors - p. Cover 6. Preface to the Second Edition - p. Cover 7. I: STRATEGIES AND TECHNIQUES - p. xviii 8. Chapter 1: Purification Strategies for Membrane Proteins - p. xviii 9. Chapter 2: Techniques and Basic Operations in Membrane Protein Purification - p. 18 10. Chapter 3: Production and Purification of Recombinant Membrane Proteins - p. 53 11. Chapter 4: SDS Electrophoresis Techniques - p. 83 12. Chapter 5: Blue Native Electrophoresis - p. 103 13. Chapter 6: Preparative Isoelectric Focusing - p. 130 14. Chapter 7: Membrane Protein Crystallization - p. 142 15. II: DISCUSSION OF SELECTED ISOLATION PROTOCOLS - p. 160 16. Chapter 8: Lipid-Dependent Inactivation and Reactivation of Bovine Complex III - p. 160 17. Chapter 9: Purification of an Affinity-Epitope Tagged G-Protein Coupled Receptor - p. 165 18. Chapter 10: Purification of NhaA Na+/H+ Antiporter of Escherichia coli for 3D or 2D Crystallization - p. 178 19. Chapter 11: Purification of the Cytochrome bc1 Complex from Yeast - p. 190 20. III: CRYSTALLIZATION OF MEMBRANE PROTEINS - p. 203 21. Chapter 12: Antibody Fragment Mediated Crystallization of Membrane Proteins - p. 203 22. Chapter 13: Crystallization of Wolinella succinogenes Quinol: Fumarate Reductase - p. 218 23. Chapter 14: Ba3 – Type Cytochrome c Oxidase from Thermus thermophilus: Purification, Crystallization and Crystal Transformation - p. 228 24. Chapter 15: Two-Dimensional Crystallization of Membrane Proteins: A Practical Guide - p. 251 25. Chapter 16: In Cubo Crystallization of Membrane Proteins - p. 284 26. Index - p. 302
This second edition of Membrane Protein Purification and Crystallization, A Practical Guide is written for bench scientists working in the fields of biochemistry, biology, and proteomic research. This guide presents isolation and crystallization techniques in a concise form, emphasizing the critical aspects unique to membrane proteins. It explains the principles of the methods and provides protocols of general use, permitting researchers and students new to this area to adapt these techniques to their particular needs. This edition is not only an update but is comprised mainly of new contributions. It is the first monograph compiling the essential approaches for membrane protein crystallization, and emphasizes recent progress in production and purification of recombinant membrane proteins.
Key features
* Provides general guidelines and strategies for isolation and crystallization of membrane proteins
* Gives detailed protocols that have wide application, and low specialized equipment needs
* Emphasizes recent progress in production and purification of recombinant membrane proteins, especially of histidine-tagged and other affinity-epitope-tagged proteins
* Summarizes recent developments of Blue-Native PAGE, a high resolution separation technique, which is independent of the use of recombinant techniques, and is especially suited for proteomic analyses of membrane protein complexes
* Gives detailed protocols for membrane protein crystallization, and describes the production and use of antibody fragments for high resolution crystallization
* Presents a comprehensive guide to 2D-crystallization of membrane proteins
1. Cover image - p. Cover 2. Title page - p. Cover 3. Table of Contents - p. Cover 4. Copyright - p. Cover 5. Contributors - p. Cover 6. Preface to the Second Edition - p. Cover 7. I: STRATEGIES AND TECHNIQUES - p. xviii 8. Chapter 1: Purification Strategies for Membrane Proteins - p. xviii 9. Chapter 2: Techniques and Basic Operations in Membrane Protein Purification - p. 18 10. Chapter 3: Production and Purification of Recombinant Membrane Proteins - p. 53 11. Chapter 4: SDS Electrophoresis Techniques - p. 83 12. Chapter 5: Blue Native Electrophoresis - p. 103 13. Chapter 6: Preparative Isoelectric Focusing - p. 130 14. Chapter 7: Membrane Protein Crystallization - p. 142 15. II: DISCUSSION OF SELECTED ISOLATION PROTOCOLS - p. 160 16. Chapter 8: Lipid-Dependent Inactivation and Reactivation of Bovine Complex III - p. 160 17. Chapter 9: Purification of an Affinity-Epitope Tagged G-Protein Coupled Receptor - p. 165 18. Chapter 10: Purification of NhaA Na+/H+ Antiporter of Escherichia coli for 3D or 2D Crystallization - p. 178 19. Chapter 11: Purification of the Cytochrome bc1 Complex from Yeast - p. 190 20. III: CRYSTALLIZATION OF MEMBRANE PROTEINS - p. 203 21. Chapter 12: Antibody Fragment Mediated Crystallization of Membrane Proteins - p. 203 22. Chapter 13: Crystallization of Wolinella succinogenes Quinol: Fumarate Reductase - p. 218 23. Chapter 14: Ba3 – Type Cytochrome c Oxidase from Thermus thermophilus: Purification, Crystallization and Crystal Transformation - p. 228 24. Chapter 15: Two-Dimensional Crystallization of Membrane Proteins: A Practical Guide - p. 251 25. Chapter 16: In Cubo Crystallization of Membrane Proteins - p. 284 26. Index - p. 302
