Heterologous Expression of Membrane Proteins

Methods and Protocols

Buch | Softcover

467 Seiten

2023 | 3rd ed. 2022
Springer-Verlag New York Inc.
978-1-0716-2370-1 (ISBN)

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This 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 protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Heterologous Expression of Membrane Proteins: Methods and Protocols, Third Edition serves as an ideal guide for scientists aiming to produce and purify functional recombinant membrane proteins for structural studies.

lt;p>1. Solubilization and Purification of a₅b₁Integrin 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 A₂G 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

Erscheinungsdatum	05.07.2023
Reihe/Serie	Methods in Molecular Biology
Zusatzinfo	78 Illustrations, color; 16 Illustrations, black and white; XVII, 467 p. 94 illus., 78 illus. in color.
Verlagsort	New York, NY
Sprache	englisch
Maße	178 x 254 mm
Themenwelt	Naturwissenschaften ► Biologie ► Biochemie
	Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie
	Naturwissenschaften ► Biologie ► Zellbiologie
	Naturwissenschaften ► Chemie
Schlagworte	Bacterial host models • Production Systems • Protein Expression • Protein Purification • Structural Studies
ISBN-10	1-0716-2370-2 / 1071623702
ISBN-13	978-1-0716-2370-1 / 9781071623701
Zustand	Neuware