Nitrogenase reaction represents a major source of the usable form of nitrogen that supports the existence of human population. As such, understanding how small building blocks are assembled into a functional nitrogenase entity is of significant relevance to human health. Using combined genetic, biochemical, spectroscopic and structural approaches, we propose to investigate how M-cluster, the active center of molybdenum nitrogenase, is assembled. Specifically, we will examine the formation of a Fe-S core of M-cluster, the maturation of Fe-S core into an M-cluster, and the transfer of M-cluster from the assembly site to its target location in great detail. Through our proposed studies, we expect to refine the biosynthetic pathway of M-cluster, which will provide crucial insights into th structural-functional relationship of nitrogenase and the general assembly mechanism of complex metal clusters in biological systems.

Public Health Relevance

Using combined genetic, biochemical, spectroscopic and structural approaches, we propose to investigate how M-cluster, the active center of molybdenum nitrogenase, is assembled. Through our proposed studies, we expect to refine the biosynthetic mechanism of M-cluster, which will provide crucial insights into the structural-functional relationship of nitrogenase and the general assembly scheme of complex metal clusters in biological systems.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM067626-10
Application #
8371547
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Anderson, Vernon
Project Start
2003-05-01
Project End
2016-11-30
Budget Start
2012-12-15
Budget End
2013-11-30
Support Year
10
Fiscal Year
2013
Total Cost
$308,127
Indirect Cost
$103,127
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 (2016) Maturation of nitrogenase cofactor-the role of a class E radical SAM methyltransferase NifB. Curr Opin Chem Biol 31:188-94
Fay, Aaron W; Blank, Michael A; Rebelein, Johannes G et al. (2016) Assembly scaffold NifEN: A structural and functional homolog of the nitrogenase catalytic component. Proc Natl Acad Sci U S A 113:9504-8
Lee, Chi Chung; Sickerman, Nathaniel S; Hu, Yilin et al. (2016) YedY: A Mononuclear Molybdenum Enzyme with a Redox-Active Ligand? Chembiochem 17:453-5
Hu, Yilin; Ribbe, Markus W (2015) Nitrogenase and homologs. J Biol Inorg Chem 20:435-45
Tanifuji, Kazuki; Lee, Chi Chung; Ohki, Yasuhiro et al. (2015) Combining a Nitrogenase Scaffold and a Synthetic Compound into an Artificial Enzyme. Angew Chem Int Ed Engl 54:14022-5
Wiig, Jared A; Hu, Yilin; Ribbe, Markus W (2015) Refining the pathway of carbide insertion into the nitrogenase M-cluster. Nat Commun 6:8034
Ribbe, Markus W (2015) Insights into the Mechanism of Carbon Monoxide Dehydrogenase at Atomic Resolution. Angew Chem Int Ed Engl 54:8337-9
Cahn, Jackson K B; Brinkmann-Chen, Sabine; Spatzal, Thomas et al. (2015) Cofactor specificity motifs and the induced fit mechanism in class I ketol-acid reductoisomerases. Biochem J 468:475-84
Lee, Chi Chung; Hu, Yilin; Ribbe, Markus W (2015) Insights into hydrocarbon formation by nitrogenase cofactor homologs. MBio 6:
Lee, Chi Chung; Fay, Aaron W; Weng, Tsu-Chien et al. (2015) Uncoupling binding of substrate CO from turnover by vanadium nitrogenase. Proc Natl Acad Sci U S A 112:13845-9

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