This book provides an overview of the different heterologous expression systems available to produce various membrane proteins for...
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This book provides an overview of the different heterologous expression systems available to produce various membrane proteins for structural analysis. Chapters examine membrane protein solubilization, purification, and instability in solution.
Membrane proteins play a key role in numerous pathologies such as cancer, cystic fibrosis, epilepsy, hyperinsulinism, and Alzheimer's disease, yet studies on these and other disorders are hampered by a lack of information about the proteins involved. In Heterologous Expression of Membrane Proteins: Methods and Protocols, expert researchers provide an overview of the different heterologous expression systems available to produce these proteins for structural analysis, helping us to understand how these proteins function, and additionally, how their functions can be modified through drug treatment. Chapters examine membrane protein solubilization, purification, and instability in solution, and investigate the strategies that helped to determine the structure of the first heterologously expressed mammalian membrane proteins. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Far-reaching and innovative, Heterologous Expression of Membrane Proteins: Methods and Protocols, is an essential guide for novice and experienced researchers alike, providing detailed methods for existing expression systems alongside cutting-edge strategies for future use in the field.
Provides descriptions of existing heterologous expression systems for membrane proteins and instructions for manipulating these systems Contains methods for the purification of membrane proteins Includes information on classical detergents and alternatives for studying membrane proteins in solution Offers methods to study and increase membrane proteins stability Includes supplementary material: sn.pub/extras Inhalt Heterologous Expression of Membrane Proteins: Methods and Protocols Isabelle Mus-Veteau, Editor Table of Contents Preface Contributors 1. Heterologous expression of membrane proteins for structural analysis Isabelle Mus-Veteau 2. Production of membrane proteins in E. coli and L. lactis Bert Poolman and Eric R. Geertsma 3. Mammalian membrane receptors expression as inclusion bodies in E. coli Bernard Mouillac and Jean-Louis Banères 4. Expression of membrane proteins at the E. coli membrane for structural studies Manuela Zoonens and Bruno Miroux 5. Membrane protein expression in Lactococcus lactis Annie Frelet-Barrand, Sylvain Boutigny, Edmund R. S. Kunji and Norbert Rolland 6. Heterologous Expression of Human Membrane Receptors in the Yeast Saccharomyces cerevisiae Olivier Joubert, Rony Nehme, Michel Bidet and Isabelle Mus-Veteau 7. Mammalian membrane proteins expression in baculovirus infected insect cells Céline Trometer and Pierre Falson 8. Expression of membrane proteins in Drosophila melanogaster S2 cells: production and analysis of a EGFP-fused G Protein-Coupled Receptor as a model Karl Brillet, Carlos A. Pereira, and Renaud Wagner 9. Membrane protein expression in the eyes of transgenic flies. Valérie Panneels and Irmgard Sinning 10. Expression of Mammalian Membrane Proteins in Mammalian Cells using Semliki Forest Virus Vectors Kenneth Lundstrom 11. Membrane Protein Expression in Cell-Free Systems Birgit Schneider, Friederike Junge, Vladimir A. Shirokov, Florian Durst, Daniel Schwarz, Volker Dötsch, Frank Bernhard 12. Practical considerations of membrane proteininstability for purification and crystallisation Christopher G. Tate 13. Membrane protein solubilization Katia Duquesne and James N. Sturgis 14. Amphipols and fluorinated surfactants: two alternatives to detergents for studying membrane proteins in vitro Cécile Breyton, Bernard Pucci and Jean-Luc Popot 15. Heterologous expression and affinity purification of eukaryotic membrane proteins in view of functional and structural studies: the example of the sarcoplasmic reticulum Ca2+-ATPase Delphine Cardi, Cédric Montigny, Bertrand Arnou, Marie Jidenko, Estelle Marchal, Marc le Maire and Christine Jaxel