Crystallography is the major method of determining structures of biological macromolecules yet crystallization techniques are still regarded as difficult to perform. This new edition of Crystallization of Nucliec Acids and Proteins: A Practical Approach continues in the vein of the first edition by providing a detailed and rational guide to producing crystals of proteins and nucleic acids of sufficient quantity and quality for diffraction studies. It has been thoroughly updated to include all the major new techniques such as the uses of molecular biology in structural biology (maximizing expression systems, sequence modifications to enable crystallization, and the introduction of anomalous scatterers); diagnostic analysis of prenucleation and nucleation by spectroscopic methods; and the two- dimensional electron crystallography of soluble proteins on planar lipid films. As well as an introduction to crystallogenesis, the other topics covered are: Handling macromolecular solutions, experimental design, seeding, proceeding from solutions to crystals Crystallization in gels Crystallization of nucleic acid complexes and membrane proteins Soaking techniques Preliminary characterization of crystals in order to tell whether they are suitable for diffraction studies. As with all Practical Approach books the protocols have been written by experienced researchers and are tried an tested methods. The underlying theory is brought together with the laboratory protocols to provide researchers with the conceptual and methodological tools necessary to exploit these powerful techniques. Crystallization of Nucliec Acids and Proteins: A Practical Approach 2e will be an invaluable manual of practical crystallization methods to researchers in molecular biology, crystallography, protein engineering, and biological chemistry.
Preface; An introduction to the crystallogenesis of biological macromolecules; Biochemical aspects and handling of macromolecular solutions and crystals; Molecular biology for structural biology; Experimental design, quantitive analysis, and the cartography of crystal growth; Methods of crystallization; Crystallization in gels and related methods; Seeding techniques; Nucleic acids and their complexes; Crystallization of membrane proteins; From solution to crystals with a physico-chemical aspect; Diagnostic of prenucleation and nucleation by spectroscopic methods and background on the physics; Two-dimensional crystallization of soluble proteins on planar lipid films; Soaking techniques; X-ray analysis; Index