Nitrogen Fixation, Softcover reprint of the original 1st ed. 2011
Methods and Protocols
Methods in Molecular Biology Series, Vol. 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.

Part I: Nitrogen Fixation

1. Historic Overview of Nitrogenase Research

            Yilin Hu and Markus W. Ribbe

2. Mechanism of Mo-Dependent Nitrogenase

            Zhi-Yong Yang, Karamatullah Danyal, and Lance C. Seefeldt

3. Assembly of Nitrogenase MoFe Protein

            Chi-Chung Lee, Aaron W. Fay, Jared A. Wiig, Markus W. Ribbe, and Yilin Hu

4. Genomic Analysis of Nitrogen Fixation

            Ina P. O’Carroll and Patricia C. Dos Santos

5. Enzymatic Systems with Homology to Nitrogenase

            Markus J. Bröcker and Jürgen Moser

Part II: Genetic and Biochemical Methods

6. Molecular Biology and Genetic Engineering in Nitrogen Fixation

            Patricia Dos Santos

7. Purification of Nitrogenase Proteins

            Jared A. Wiig, Chi-Chung Lee, Aaron W. Fay, Yilin Hu, and Markus W. Ribbe

8. Assays of Nitrogenase Reaction Products

            William E. Newton and Michael J. Dilworth

9. Methods for Nitrogenase-Like Dark Operative Protochlorophyllide Oxidoreductase

            Markus J. Bröcker and Jürgen Moser

Part III: Spectroscopic Methods

10. X-Ray Crystallography

            Lauren E. Roth and F. Akif Tezcan

11. X-Ray Absorption Spectroscopy

            Serena DeBeer

12. Small Angle X-Ray Scattering Spectroscopy

            David W. Mulder and John W. Peters

13. Electron Paramagnetic Resonance Spectroscopy

            Karamatullah Danyal, Zhi-Yong Yang, and Lance C. Seefeldt

14. Magnetic Circular Dichroism Spectroscopy

            Brian J. Hales

15. Mössbauer Spectroscopy

            Boi Hanh Huynh

Part IV: Chemical Methods

16. Protocols for Cofactor Isoation of Nitrogenase

            Aaron W. Fay, Chi-Chung Lee, Jared A. Wiig, Yilin Hu, and Markus W. Ribbe

17. Techniques for Functional and Structural Modeling of Nitrogenase

            Patrick L. Holland

Part V: Theoretical Calculations

18. Nitrogenase Structure and Function Relationships by Density Functional Theory

            Travis V. Harris and Robert K. Szilagyi

19. Modeling the MoFe Nitrogenase System with Broken Symmetry Density Functional Theory

            Gregory M. Sandala and Louis Noodleman