Heterologous Expression of Membrane Proteins: Methods and Protocols

by Isabelle Mus-Veteau (Editor)

Overview

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.

Product Details

ISBN-13:	9781071623701
Publisher:	Springer US
Publication date:	07/02/2023
Series:	Methods in Molecular Biology , #2507
Edition description:	3rd ed. 2022
Pages:	467
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Preface v

Contributors ix

1 Heterologous Expression of Membrane Proteins for Structural Analysis Isabelle Mus-Veteau 1

2 Production of Membrane Proteins in Escherichia coli and Lactococcus lactis Eric R. Geertsma Bert Poolman 17

3 Mammalian Membrane Receptors Expression as Inclusion Bodies in Escherichia coli Bernard Mouillac Jean-Louis Banères 39

4 Expression of Membrane Proteins at the Escherichia coli Membrane for Structural Studies Manuela Zoonens Bruno Miroux 49

5 Membrane Protein Expression in Lactococcus lactis Annie Frelet-Barrand Sylvain Boutigny Edmund R.S. Kunji Norbert Rolland 67

6 Heterologous Expression of Human Membrane Receptors in the Yeast Saccharomyces cerevisiae Olivier Joubert Rony Nehmé Michel Bidet Isabelle Mus-Veteau 87

7 Mammalian Membrane Protein Expression in Baculovirus-Infected Insect Cells Céline Trometer Pierre Falson 105

8 Expression of membrane proteins in Drosophila melanogaster S2 Cells: Production and Analysis of a EGFP-Fused G Protein-Coupled Receptor as a Model Karl Brillet Carlos A. Pereira Renaud Wagner 119

9 Membrane Protein Expression in the Eyes of Transgenic Flies Valérie Panneels Irmgard Sinning 135

10 Expression of Mammalian Membrane Proteins in Mammalian Cells Using Semliki Forest Virus Vectors Kenneth Lundstrom 149

11 Membrane Protein Expression in Cell-Free Systems Birgit Schneider Friederike Junge Vladimir A. Shirokov Florian Durst Daniel Schwarz Volker Dötsch Frank Bernhard 165

12 Practical Considerations of Membrane Protein Instability during Purification and Crystallisation Christopher G. Tate 187

13 Membrane Protein Solubilization Katia Duquesne James N. Sturgis 205

14 Amphipols and Fluorinated Surfactants: Two Alternatives to Detergents for Studying Membrane Proteins In vitro Cécile Breyton Bernard Pucci Jean-Luc Popot 219

15 Heterologous Expression and Affinity Purification of Eukaryotic Membrane Proteins in View of Functional and Structural Studies: The Example of the Sarcoplasmic Reticulum Ca²⁺-ATPase Delphine Cardi Cédric Montigny Bertrand Arnou Marie Jidenko Estelle Marchal Marc Le Maire Christine Jaxel 247

Index 269

Editorial Reviews

Reviewer: Omer M. Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: This well-illustrated book provides an overview of different heterologous expression systems available to produce membrane proteins for structural analysis to facilitate understanding of how these proteins function and how their functions can be modified by drug treatment.
Purpose: It is intended as an essential guide for novice and experienced researchers.
Audience: Written for students and experienced researchers, the book also will be useful for clinicians interested in the care of patients with cancer, epilepsy, hyperinsulinism, hypertension, cystic fibrosis, and Alzheimer's disease.
Features: Each of the 15 chapters is written by an expert in the topic. Chapters deal with heterologous expression of membrane proteins for structural analysis, production of membrane proteins in E. coli and L. lactis, membrane protein solubilization, purification, and instability in solution. Each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a notes section with tips on troubleshooting and avoiding known pitfalls. The book has several black-and-white and color illustrations.
Assessment: Part of the highly successful Methods in Molecular Biology series, this book provides valuable information on membrane proteins for novice and experienced researchers. Clinicians will find it extremely useful for developing methods and protocols for research.

Doody's Review Service

From the reviews:

“Well-illustrated book provides an overview of different heterologous expression systems available to produce membrane proteins for structural analysis to facilitate understanding of how these proteins function and how their functions can be modified by drug treatment. … Written for students and experienced researchers, the book also will be useful for clinicians interested in the care of patients with cancer … . provides valuable information on membrane proteins for novice and experienced researchers. Clinicians will find it extremely useful for developing methods and prools for research.” (Omer Iqbal, Doody’s Review Service, July, 2010)

From the Publisher