(Ebook PDF) Heterologous Expression of Membrane Proteins Methods and Protocols 1st Edition by Isabelle Mus Veteau 1493961322 9781493961320 Full chapter

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ISBN 10:         1493961322
ISBN 13: 978-1493961320
Author:          Isabelle Mus Veteau

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.

Heterologous Expression of Membrane Proteins Methods and Protocols 1st  Table of contents:

Part I: Basic Methods and Protocols

1. Introduction to Membrane Protein Expression

   • Overview of membrane proteins and the challenges associated with their expression.

2. Selection of Expression Systems for Membrane Proteins

   • Discussing different expression systems (e.g., bacterial, yeast, mammalian) used for membrane proteins.

3. Expression of Membrane Proteins in Escherichia coli

   • Step-by-step protocols for expressing membrane proteins in E. coli.

4. Expression in Yeast Systems

   • Techniques for the heterologous expression of membrane proteins in yeast.

5. Expression in Insect Cells

   • Methods for utilizing insect cells for membrane protein expression.

6. Expression in Mammalian Cells

   • Protocols for expressing membrane proteins in mammalian cells.

Part II: Protein Isolation and Purification
7. Isolation of Membrane Fractions

• Detailed methods for isolating membrane proteins from cells.

8. Solubilization of Membrane Proteins

   • Techniques for solubilizing membrane proteins without losing their functionality.

9. Purification of Membrane Proteins

   • Protocols for purifying membrane proteins from solubilized membranes.

Part III: Characterization and Functional Assays
10. Determining Protein Quality and Integrity
– Methods for assessing the integrity of expressed membrane proteins.

11. Functional Assays for Membrane Proteins

    • Different assays to test the functional activity of membrane proteins post-expression.

12. Structural Characterization of Membrane Proteins

    • Techniques for determining the structure of membrane proteins, including cryo-EM and X-ray crystallography.

Part IV: Advanced Techniques and Applications
13. Incorporation of Membrane Proteins into Liposomes
– Procedures for reconstituting membrane proteins into lipid bilayers for functional studies.

14. Studying Membrane Protein Interactions

    • Methods for identifying and analyzing protein-protein interactions in membrane proteins.

15. Applications of Membrane Protein Expression in Drug Discovery

    • The role of heterologous expression of membrane proteins in drug development and screening.

Part V: Troubleshooting and Optimization
16. Troubleshooting Common Problems in Membrane Protein Expression
– Solutions to common issues faced during the expression and purification of membrane proteins.

17. Optimizing Membrane Protein Expression and Purification
    • Strategies for improving the yield and functionality of membrane proteins.

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