Ribbe M.W. (Ed.) Nitrogen Fixation: Methods and Protocols

Humana Press, 2011. - 337 p. - Methods in Molecular Biology, Volume
766. - Few problems in protein biochemistry have proven to be as challenging and recalcitrant as the molecular description of nitrogenase, the catalyst of one of the most remarkable chemical transformations in biological systems: the nucleotide-dependent reduction of atmospheric dinitrogen to bioavailable ammonia. In Nitrogen Fixation: Methods and Protocols, recognized experts in the field provide an up-to-date, in-depth overview of the methods that have been applied to studying the nitrogenase at a molecular level, ranging from genetic, biochemical, spectroscopic, and chemical methods to theoretical calculations. In addition, techniques used to study an enzyme system that is homologous to nitrogenase are described in this book. Written in the highly successful Methods in Molecular Biology series format, methods chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Nitrogen Fixation: Methods and Protocols will be useful for anyone interested in nitrogenase research and willing to venture further toward addressing the remaining mechanistic and biosynthetic questions of this fascinating enzyme system.Contents
Nitrogen Fixation
Historic Overview of Nitrogenase Research
Mechanism of Mo-Dependent Nitrogenase
Assembly of Nitrogenase MoFe Protein
Genomic Analysis of Nitrogen Fixation
Enzymatic Systems with Homology to Nitrogenase
Genetic and Biochemical Methods
Molecular Biology and Genetic Engineering in Nitrogen Fixation
Purification of Nitrogenase Proteins
Assays of Nitrogenase Reaction Products
Methods for Nitrogenase-Like Dark Operative Prohlorophyllide Oxidoreductase
Spectroscopic Methods
X-Ray Crystallography
X-Ray Absorption Spectroscopy
Small Angle X-Ray Scattering Spectroscopy
Electron Paramagnetic Resonance Spectroscopy
Magnetic Circular Dichroism Spectroscopy
Mössbauer Spectroscopy
Chemical Methods
Prools for Cofactor Isoation of Nitrogenase
Techniques for Functional and Structural Modeling of Nitrogenase
Theoretical Calculations
Nitrogenase Structure and Function Relationships by Density Functional Theory
Modeling the MoFe Nitrogenase System with Broken Symmetry Density Functional Theory