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 laboratoryprotocols, 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.
Reihe
Auflage
Sprache
Verlagsort
Verlagsgruppe
Illustrationen
78
16 s/w Abbildungen, 78 farbige Abbildungen
XVII, 467 p. 94 illus., 78 illus. in color.
ISBN-13
978-1-0716-2368-8 (9781071623688)
DOI
10.1007/978-1-0716-2368-8
Schweitzer Klassifikation
Solubilization and Purification of a
5
b
1
Integrin from Rat Liver for Reconstitution into Nanodiscs.- Membrane Proteins Production in
Escherichia coli
: Protocols and Rules.- Functional Overexpression of Membrane Proteins in E. coli: The Good, the Bad, and the Ugly.- Heterologous Expression of Membrane Proteins in E. coli.- Overexpression of the ABC Transporter BmrA within Intracellular Caveolæ in
Escherichia coli
.- Production of Membrane Proteins in
Pseudomonas stutzeri
.- Overproduction of Membrane-Associated, and Integrated, Proteins Using
Saccharomyces cerevisiae
.- Purification of Membrane Proteins Overexpressed in
Saccharomyces cerevisiae
.- Production and Purification of a GFP-Tagged ABC Transporter
CaCdr1p
.- Membrane Protein Production in the Yeast,
P. pastoris
.- Production and Preparation of Isotopically-Labelled Human Membrane Proteins in
Pichia pastoris
for Fast-MAS-NMR Analyses.- Membrane Protein Production in Insect Cells.- Thromboxane A
2
G Protein-Coupled Receptor Production and Crystallization for Structure Studies.- Production of Human ABC Transporters and Oligosaccharyltransferase Complexes for Structural Studies.- The Heterologous Expression and Purification of GPCRs.- Expression and Purification of the Human Thyroid-Stimulating Hormone Receptor.- Optimization of Recombinant GPCR Proteins for Biophysical and Structural Studies Using Virus-Like Particles.- Peptide Tags and Domains for Expression and Detection of Mammalian Membrane Proteins at the Cell Surface.- Tailor the Purification and Analysis of Membrane Proteins with Modular Detergents.- Detergent Alternatives: Membrane Protein Purification Using Synthetic Nanodisc Polymers.- Detergent-Free Membrane Protein Purification Using SMA Polymer.- Cell-Free Expression of GPCRs into Nanomembranes for Functional and Structural Studies.- Cell-Free Expression of Proton-Coupled Folate Transporter in the Presence of Nanodiscs.- Microscale Thermophoresis to Evaluate the Functionality of Heterologously Overexpressed Membrane Proteins in Membrane Preparations.
