Expression of Membrane Proteins -- Expression and Purification of the Amphipathic Form of Rabbit Cytochrome b5 in Escherichia coli -- Dihydroorotate Dehydrogenase of Escherichia coli -- A General Approach for Heterologous Membrane Protein Expression in Escherichia coli -- Expression of Membrane-Bound Iron-Sulfur Proteins -- Heterologous Expression of Human Scavenger Receptor Class B Type I (SR-BI) in Pichia pastoris -- Expression of Oligomeric Amiloride-Sensitive Epithelial Sodium Channel in Sf9 Insect Cells -- Functional Expression of His-Tagged Rhodopsin in Sf9 Insect Cells -- Detergent Selection in Membrane Protein Purification -- Preparation of Glycerol Facilitator for Protein Structure and Folding Studies in Solution -- Solubilization of Chemokine Receptors from Cell Membranes -- A Systematic Approach for the Solubilization of the Integral Membrane Protein Lysophospholipid -- Membrane Protein Purification -- Purification of Omp50, a Minor Porin of Campylobacter jejuni -- Purification of Porins from Mycobacterium smegmatis -- Isolation of the Melanocortin 5 Receptor -- Purification of Mammalian Serine Palmitoyltransferase, a Hetero-Subunit Enzyme for Sphingolipid Biosynthesis, by Affinity-Peptide Chromatography -- Purification of Phosphatidylglycerophosphate Synthase from Cultured Mammalian Cells -- Purification of Pancreastatin Receptor from Rat Liver Membranes -- Purification, Reconstitution, and Functional Characterization of Zinc Transporter from Rat Renal Brush Border Membranes -- Isolation of Lipid Raft-Associated Proteolipids -- Purification of Membrane-Bound Catechol-O-Methyltransferase -- Purification and Characterization of Transporter Proteins from Human Erythrocyte Membrane -- Purification of the Human Erythrocyte PS-Stimulated Mg2+-ATPase -- Reconstitution and Assay of Biogenic Membrane-Derived Phospholipid Flippase Activity in Proteoliposomes -- Expression, Purification, and Reconstitution of Rat Liver Carnitine Palmitoyltransferase I -- Structural Analysis of Membrane Proteins -- Crystallization in Lipidic Cubic Phases -- Optical Biosensor Assay Using Retroviral Receptor Pseudotypes.

Recent developments in molecular biology and biochemistry have made it possible to determine the three-dimensional structure of membrane proteins and so come to a full understanding of their function. In Membrane Protein Protocols: Expression, Purification, and Characterization, researchers at major universities and research centers around the world describe in detail the key techniques that have proven successful in the study of receptors and transport proteins. The book provides examples of how different membrane proteins can be overexpressed in both prokaryotic and eukaryotic expression systems, how natural and overexpressed proteins can be solubilized from their host membranes, and how the solubilized protein can be purified in active form. Each protocol contains step-by-step instructions to ensure success, troubleshooting advice, lists of reagents, and tips on avoiding pitfalls. Timely and highly practical, Membrane Protein Protocols: Expression, Purification, and Characterization offers investigators proven methods to determine the three-dimensional structure of membrane receptors and transport proteins, as well as proven practical help in the discovery of new clinical targets for the design of novel pharmaceutical agents useful in treating cancer, bacterial and viral infection, and many metabolic diseases.