Nitrogenase is a complex metalloenzyme that catalyzes one of the most remarkable chemical transformations in biological systems: the nucleotide-dependent reduction of atmospheric dinitrogen to bioavailable ammonia. There is an outstanding interest in decoding the assembly mechanism of nitrogenase, not only because of its significant relevance to nitrogenase-specific reactions, but also because of its tremendous implication for general metalloprotein biogenesis. Here we propose to greatly expand our understanding of the nitrogenase assembly process by combined genetic, biochemical, spectroscopic and structural approaches. The MoFe protein of the molybdenum nitrogenase of Azotobacter vinelandii will be the focus of the proposed investigation. Through our proposed studies, we expect to further refine the biosynthetic pathways of MoFe protein and its associated clusters, a continuous effort with an ultimate goal to unravel the molecular mechanism of nitrogenase assembly. Furthermore, using nitrogenase as a model system, we also hope to provide general themes of biological metal cluster assembly, which may, in the long run, prove instrumental in developing successful strategies for the chemical synthesis of complex metalloclusters. .

Public Health Relevance

Nitrogenase catalyzes the transformation of atmospheric dinitrogen to bioavailable ammonia, thereby supplying the reduced nitrogen, an essential component of nucleic acids and proteins, for the existence of human population. This particular proposal deals with the assembly of nitrogenase MoFe protein and its associated metalloclusters and the outcome of these studies will hopefully provide general themes of biological metal cluster assembly and facilitate future development of cost-efficient strategies for the chemical synthesis of catalytically active metalloclusters. As such, our proposed research may have additional economic value in addition to its intrinsic relevancy to human health.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM067626-09
Application #
8197578
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Anderson, Vernon
Project Start
2003-05-01
Project End
2012-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
9
Fiscal Year
2012
Total Cost
$289,382
Indirect Cost
$93,362
Name
University of California Irvine
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Hu, Yilin; Ribbe, Markus W (2015) Nitrogenase and homologs. J Biol Inorg Chem 20:435-45
Lee, Chi Chung; Hu, Yilin; Ribbe, Markus W (2015) Catalytic reduction of CN-, CO, and CO2 by nitrogenase cofactors in lanthanide-driven reactions. Angew Chem Int Ed Engl 54:1219-22
Rebelein, Johannes G; Hu, Yilin; Ribbe, Markus W (2014) Differential reduction of CO? by molybdenum and vanadium nitrogenases. Angew Chem Int Ed Engl 53:11543-6
Hu, Yilin; Ribbe, Markus W (2014) A journey into the active center of nitrogenase. J Biol Inorg Chem 19:731-6
Ribbe, Markus W; Hu, Yilin; Hodgson, Keith O et al. (2014) Biosynthesis of nitrogenase metalloclusters. Chem Rev 114:4063-80
Rupnik, Kresimir; Lee, Chi Chung; Wiig, Jared A et al. (2014) Nonenzymatic synthesis of the P-cluster in the nitrogenase MoFe protein: evidence of the involvement of all-ferrous [Fe4S4](0) intermediates. Biochemistry 53:1108-16
Hu, Yilin; Ribbe, Markus W (2013) Biosynthesis of the iron-molybdenum cofactor of nitrogenase. J Biol Chem 288:13173-7
Lancaster, Kyle M; Hu, Yilin; Bergmann, Uwe et al. (2013) X-ray spectroscopic observation of an interstitial carbide in NifEN-bound FeMoco precursor. J Am Chem Soc 135:610-2
Wiig, Jared A; Lee, Chi Chung; Hu, Yilin et al. (2013) Tracing the interstitial carbide of the nitrogenase cofactor during substrate turnover. J Am Chem Soc 135:4982-3
Hu, Yilin; Ribbe, Markus W (2013) Nitrogenase assembly. Biochim Biophys Acta 1827:1112-22

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