Membrane Protein Purification and Crystallization: A Practical Guide

This second edition of Membrane Protein Purification and Crystallization, A Practical Guide is written for bench scientists working in the fields of biochemistry, biology, and proteomic research. This guide presents isolation and crystallization techniques in a concise form, emphasizing the critical aspects unique to membrane proteins. It explains the principles of the methods and provides protocols of general use, permitting researchers and students new to this area to adapt these techniques to their particular needs. This edition is not only an update but is comprised mainly of new contributions. It is the first monograph compiling the essential approaches for membrane protein crystallization, and emphasizes recent progress in production and purification of recombinant membrane proteins.

Provides general guidelines and strategies for isolation and crystallization of membrane proteinsGives detailed protocols that have wide application, and low specialized equipment needsEmphasizes recent progress in production and purification of recombinant membrane proteins, especially of histidine-tagged and other affinity-epitope-tagged proteins Summarizes recent developments of Blue-Native PAGE, a high resolution separation technique, which is independent of the use of recombinant techniques, and is especially suited for proteomic analyses of membrane protein complexesGives detailed protocols for membrane protein crystallization, and describes the production and use of antibody fragments for high resolution crystallizationPresents a comprehensive guide to 2D-crystallization of membrane proteins

I. Strategies and Techniques Purification Strategies for Membrane Proteins Chromatographic Techniques and Basic Operations in Membrane Protein Purification Production and purification of recombinant membrane proteins SDS Electrophoresis Techniques Blue-Native Electrophoresis Preparative Isoelectric Focussing Membrane protein crystallization

II. Discussion of Selected Isolation Protocols Lipid Dependent Inactivation and Reactivation of Bovine complex III Purification of affinity-epitope tagged G-protein coupled receptors Purification of NhaA, the Na+/H+ Antiporter of Escherichia coli for 3D or 2D Crystallization Purification of the yeast cytochrome bc1 complex

III. Crystallization of Membrane Proteins Antibody fragment-mediated crystallization of membrane proteins Crystallization of Wolinella succinogenes quinol:fumarate reductase Ba3-type cytochrome-c oxidase from Thermus thermophilus purification, crystallization, and crystal transformation Two-dimensional crystallization of membrane proteins: a practical guide In cubo crystallization of membrane proteins