Advanced Methods in Structural Biology

This book provides practical information on a whole set of protein experiments for advanced structural biology, such as X-ray crystallography, NMR, electron microscopy, advanced mass spectroscopy, and surface plasmon resonance, as well as a wide variety of expression systems including eukaryotic and in vitro expression. In the past decade, structural genomics studies have pushed forward the development of automated methods in the field of structural biology, however there is an increasing need for the structural analysis of difficult targets, such as large protein complexes and membrane proteins, which are hard to achieve using conventional automated methods, and require knowledge that goes beyond standard protein chemistry protocols. To address these problems and to help researchers develop novel methods, this volume provides examples of the development of new protein investigation methods and their theoretical background. This book particularly appeals to graduate students, postdoctoral researchers, young investigators wishing to gain a better understanding of the theory behind experiments, and those seeking further advanced, practical structural biology methods.

Part 1. Protein expression. - 1 Expression
in Bacteria and Refolding. - 2 Expression of proteins in insect and mammalian
cells. - 3 Application of MultiBac system to large complexes. - 4 Purification
using affinity-tag technology. - 5 Cell-Free Protein Production for Structural
Biology. - Part 2. Purification and crystallization of membrane proteins. - 6 Overview
of membrane protein purification and crystallization. - 7 Cell-free synthesis of
membrane proteins. - Part 3. Crystallization and Crystal Engineering. - 8 Screening
of cryoprotectants and the multistep soaking method. - 9 Protein modification
for crystallization. - Part 4. Interaction analysis. - 10 Analytical
ultracentrifugation. - 11 Mass spectrometry. - 12 Frontal gel filtration. - 13 Surface
Plasmon Resonance. - Part 5. Advanced methods for structural analyses. - 14 Structural
biology with micro-focus beam-lines. - 15 Structural biology and electron
microscopy. - 16 Structure determination software for macromolecular X-ray
crystallography. - 17 NMR Structural Biology using Paramagnetic Lanthanide Probe.