Heterologous Expression of Membrane Proteins

ISBN-13: 9781071623701 / Angielski / Miękka / 2023

Opis

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.

Spis treści

1. 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