Main description:

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 (TM) 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:

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. Broecker and Jurgen 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. Broecker and Jurgen 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. Moessbauer 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