1. Cloning and Multi-Subunit Expression of Mitochondrial Membrane Protein Complexes in Saccharomyces cerevisiae
Porsha L.R. Shaw, Kathryn A. Diederichs, Ashley Pitt, Sarah E. Rollauer, and Susan K. Buchanan
2. Membrane Protein Production in Escherichia coli
Benjamin C. McIlwain and Ali A. Kermani
3. Membrane Protein Production in Lactococcus Lactis for Structural Studies
Chloe Martens
4. Expression and Purification of Membrane Proteins in Saccharomyces cerevisiae
Martin S. King and Edmund R.S. Kunji
5. Membrane Protein Expression in Insect Cells Using the Baculovirus Expression Vector System
Jacques Boivineau, Matthias Haffke, and Veli-Pekka Jaakola
6. Membrane Extracts from Plant Tissues
Uwe Bodensohn, Christoph Ruland, Roman Ladig, and Enrico Schleiff
7. Membrane Protein Solubilization and Quality Control: An Example of a Primary Active Transporter
Charlott Stock and Inga Hänelt
8. GPCR Solubilization and Quality Control
Tamara Miljus, David A Sykes, Clare Harwood, Ziva Vuckovic, and Dmitry B. Veprintsev
9. Affinity Purification of Membrane Proteins
Elisabeth Graeber and Volodymyr M. Korkhov
10. Purification of Membrane Proteins by Affinity Chromatography with On-Column Protease Cleavage
Stephan Hirschi and Dimitrios Fotiadis
11. Biotinylation of Membrane Proteins for Binder Selections
Benedikt T. Kuhn, Iwan Zimmermann, Pascal Egloff, Lea M. Hürlimann, Cedric A.J. Hutter, Christian Miscenic, Roger J.P. Dawson, Markus A. Seeger, and Eric R. Geertsma
12. Production and Application of Nanobodies for Membrane Protein Structural Biology
Janine Denise Brunner and Stephan Schenck
13. Identifying Conformation-Selective Heavy-Chain-Only Antibodies against Membrane Proteins by a Thermal-Shift Scintillation Proximity Assay
Peter Stohler and Roger J.P. Dawson
14. Reconstitution of Membrane Proteins into Platforms Suitable for Biophysical and Structural Analyses
Philipp A.M. Schmidpeter, Nattakan Sukomon, and Crina M. Nimigean
15. Reconstitution of Proteoliposomes for Phospholipid Scrambling and Non-Selective Channel Assays
Maria E. Falzone and Alessio Accardi
16. Membrane Protein Cryo-EM: Cryo-Grid Optimization and Data Collection with Protein in Detergent
Magnus Bloch, Mònica Santiveri, and Nicholas M.I. Taylor
17. Single-Particle Cryo-EM of Membrane Proteins in Lipid Nanodiscs
Valeria Kalienkova, Carolina Alvadia, Vanessa Clerico Mosina, and Cristina Paulino
18. Fast Small-Scale Membrane Protein Purification and Grids Preparation for Single Particle Electron Microscopy
Natalie Bärland and Camilo Perez
19. Stabilization and Crystallization of a Membrane Protein Involved in Lipids Transport
Bing Zhang and Camilo Perez
20. In Meso In Situ Serial X-ray Crystallography (IMISX): A Protocol for Membrane Protein Structure Determination at the Swiss Light Source
Chia-Ying Huang, Vincent Olieric, Martin Caffrey, and Meitian Wang
21. Membrane Protein Preparation for Serial Crystallography Using High Viscosity Injectors: Rhodopsin as an Example
Tobias Weinert and Valérie Panneels
22. Hydrogen/Deuterium Exchange Mass Spectrometry for the Structural Analysis of Detergent-Solubilized Membrane Proteins
Darragh P. O’Brien, Véronique Hourdel, Alexandre Chenal, and Sébastien Brier
23. Mechanical Unfolding and Refolding of Single Membrane Proteins by Atomic Force Microscopy
Noah Ritzmann and Johannes Thoma
24. Sample Preparation and Technical Setup for NMR Spectroscopy with Integral Membrane Proteins
Hundeep Kaur, Anne Grahl, Jean-Baptiste Hartmann, and Sebastian Hiller
25. Exploring Lipid- and Membrane Protein-Dynamics Using Lipid-Bilayer Nanodiscs and Solution-State NMR Spectroscopy
Stefan Bibow
This book collects up-to-date advanced protocols and advice from leading experts in the area of membrane protein biology that can be applied to structural and functional studies of any membrane protein system. The contents explore methods for cloning and expression of membrane proteins and membrane protein complexes in prokaryotic and eukaryotic systems, approaches for protein purification, nanobody applications, as well as biophysical characterization and much more. Written for the highly successful Methods in Molecular Biology series, 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 thorough, Expression, Purification, and Structure Biology of Membrane Proteins serves to guide and encourage young researchers and newcomers to the field to tackle bold new studies on membrane proteins.
Chapter 11 is available open access under a CC-BY 4.0 license via link.springer.com.