Description
Book SynopsisThis detailed volume explores protocols for the production of membrane proteins in a panel of heterologous organisms for structural studies. Beginning with techniques using
E. coli as a host for the overproduction and purification of membrane proteins, the book continues with chapters covering mammalian membrane protein production in yeast, insect cells, mammalian cells, as well as using virus like particles and acellular systems. Additionally, new detergents and alternatives to detergents allowing membrane protein purification for structural analyses are described. The book closes with a chapter exploring the use of microscale thermophoresis (MST) to evaluate the binding activity of heterologously expressed proteins directly in crude membrane extracts. 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
Table of Contents1. Solubilization and Purification of a5b1 Integrin from Rat Liver for Reconstitution into Nanodiscs
Estelle Dransart, Aurélie Di Cicco, Ahmed El Marjou, Daniel Lévy, Staffan Johansson, Ludger Johannes, and Massiullah Shafaq-Zadah
2. Membrane Proteins Production in Escherichia coli: Protocols and Rules
Jordi Royes, Pauline Talbot, Christel Le Bon, Karine Moncoq, Marc Uzan, Francesca Zito, and Bruno Miroux
3. Functional Overexpression of Membrane Proteins in E. coli: The Good, the Bad, and the Ugly
Margot Di Cesare, Aissatou Maty Diagne, Benjamin Bourgey, Jean-Michel Jault, and Cédric Orelle
4. Heterologous Expression of Membrane Proteins in E. coli
Peer Depping, María Monserrat Román Lara, Athanasios Kesidis, Roslyn M. Bill, Alice J. Rothnie, Douglas F. Browning, and Alan D. Goddard
5. Overexpression of the ABC Transporter BmrA within Intracellular Caveolæ in Escherichia coli
Alexia Gobet, Christine Jaxel, Sandrine Magnard, Manuel Garrigos, Stéphane Orlowski, Nadège Jamin, Pierre Falson, and Vincent Chaptal
6. Production of Membrane Proteins in Pseudomonas stutzeri
Hao Xie, Cornelia Muenke, Manuel Sommer, Sabine Buschmann, and Hartmut Michel
7. Overproduction of Membrane-Associated, and Integrated, Proteins Using Saccharomyces cerevisiae
Landon Haslem, Marina Brown, Xin A. Zhang, Jennifer M. Hays, and Franklin A. Hays
8. Purification of Membrane Proteins Overexpressed in Saccharomyces cerevisiae
Landon Haslem, Marina Brown, Xin A. Zhang, Jennifer M. Hays, and Franklin A. Hays
9. Production and Purification of a GFP-Tagged ABC Transporter CaCdr1p
Jorgaq Pata, Alexis Moreno, Sandrine Magnard, Atanu Banerjee, Rajendra Prasad, and Pierre Falson
10. Membrane Protein Production in the Yeast, P. pastoris
Hoor Ayub, Michelle Clare, Luke Broadbent, John Simms, Alan D. Goddard, Alice J. Rothnie, and Roslyn M. Bill
11. Production and Preparation of Isotopically-Labelled Human Membrane Proteins in Pichia pastoris for Fast-MAS-NMR Analyses
Lina Barret, Tobias Schubeis, Valérie Kugler, Lucile Guyot, Guido Pintacuda, and Renaud Wagner
12. Membrane Protein Production in Insect Cells
Afroditi Vaitsopoulou, Peer Depping, Roslyn M. Bill, Alan D. Goddard, and Alice J. Rothnie
13. Thromboxane A2 G Protein-Coupled Receptor Production and Crystallization for Structure Studies
Pawel Krawinski and Martin Caffrey
14. Production of Human ABC Transporters and Oligosaccharyltransferase Complexes for Structural Studies
Ana S. Ramírez, Kamil Nosol, and Kaspar P. Locher
15. The Heterologous Expression and Purification of GPCRs
Xiaoting Li, Ling Shen, Junlin Liu, Xiaoyan Liu, Zhi-Jie Liu, and Tian Hua
16. Expression and Purification of the Human Thyroid-Stimulating Hormone Receptor
Lukas Helfinger and Christopher G. Tate
17. Optimization of Recombinant GPCR Proteins for Biophysical and Structural Studies Using Virus-Like Particles
Kathleen Aertgeerts, Thao T. Ho, and Yingzhou G. Yan
18. Peptide Tags and Domains for Expression and Detection of Mammalian Membrane Proteins at the Cell Surface
Valeria Jaramillo-Martinez, Vadivel Ganapathy, and Ina L. Urbatsch
19. Tailor the Purification and Analysis of Membrane Proteins with Modular Detergents
Leonhard H. Urner
20. Detergent Alternatives: Membrane Protein Purification Using Synthetic Nanodisc Polymers
Valentina S. Dimitrova, Saemee Song, Alexandra Karagiaridi, Anika Marand, and Heather W. Pinkett
21. Detergent-Free Membrane Protein Purification Using SMA Polymer
Luke Broadbent, Peer Depping, Alexis Lodé, Afroditi Vaitsopoulou, David Hardy, Hoor Ayub, James Mitchell-White, Ian D. Kerr, Alan D. Goddard, Roslyn M. Bill, and Alice J. Rothnie
22. Cell-Free Expression of GPCRs into Nanomembranes for Functional and Structural Studies
Simon Umbach, Volker Dötsch, and Frank Bernhard
23. Cell-Free Expression of Proton-Coupled Folate Transporter in the Presence of Nanodiscs
Hoa Quynh Do and Michaela Jansen
24. Microscale Thermophoresis to Evaluate the Functionality of Heterologously Overexpressed Membrane Proteins in Membrane Preparations
Sandra Kovachka, Pierre Soule, and Isabelle Mus-Veteau
