The long-term goal of this research is to elucidate the reaction mechanisms of NO reduction that occur at heme and non-heme diiron centers of bacterial metalloenzymes. Our studies will focus on two enzymatic systems and related bioengineered or synthetic models: 1) NO-detoxifying flavodiiron proteins (FDPs) and 2) denitrifying NO reductases (denNORs). All of the proteins within these two families are known to reduce NO to the unreactive N2O product, but they do so with wide variation in efficiency and protein matrix structure. Although several of these proteins have been characterized by X-ray crystallography, the initial steps of NO binding, iron-nitrosyl reduction, and how these catalytic events differ between systems are not well understood. The coupling of resonance Raman, FTIR, and EPR spectroscopies with rapid-freeze-quench analyses provides unique capabilities to define NO-binding geometries at diiron clusters and to follow the N-N bond formation, N-O bond cleavage, and protonation steps that must take place to convert two NO molecules to N2O and H2O. Studying a diverse group of native enzymes and models will allow us to compare and contrast structural information on iron-nitrosyl intermediates and the efficiency of the reductive and proton transfer steps of this reaction. Public health relevance: A better understanding of microbial NO reductases is highly desirable since these enzymatic reactions lead to microorganisms'resistance to the mammalian immune response. Furthermore, there are no human orthologs to these microbial enzymes;they represent potential targets for new drugs.

Public Health Relevance

A better understanding of microbial NO reductases is highly desirable since these enzymatic reactions lead to microorganisms'resistance to the mammalian immune response. Furthermore, there are no human orthologs to these microbial enzymes;they represent potential targets for new drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM074785-07
Application #
8474781
Study Section
Special Emphasis Panel (ZRG1-BCMB-B (02))
Program Officer
Anderson, Vernon
Project Start
2005-09-01
Project End
2016-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
7
Fiscal Year
2013
Total Cost
$276,693
Indirect Cost
$93,343
Name
Oregon Health and Science University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Bhagi-Damodaran, Ambika; Reed, Julian H; Zhu, Qianhong et al. (2018) Heme redox potentials hold the key to reactivity differences between nitric oxide reductase and heme-copper oxidase. Proc Natl Acad Sci U S A 115:6195-6200
Sabuncu, Sinan; Reed, Julian H; Lu, Yi et al. (2018) Nitric oxide reductase activity in heme-nonheme binuclear engineered myoglobins through a one-electron reduction cycle. J Am Chem Soc :
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Bhagi-Damodaran, Ambika; Michael, Matthew A; Zhu, Qianhong et al. (2017) Why copper is preferred over iron for oxygen activation and reduction in haem-copper oxidases. Nat Chem 9:257-263
McQuilken, Alison C; Matsumura, Hirotoshi; Dürr, Maximilian et al. (2016) Photoinitiated Reactivity of a Thiolate-Ligated, Spin-Crossover Nonheme {FeNO}(7) Complex with Dioxygen. J Am Chem Soc 138:3107-17
Basudhar, Debashree; Madrona, Yarrow; Yukl, Erik T et al. (2016) Distal Hydrogen-bonding Interactions in Ligand Sensing and Signaling by Mycobacterium tuberculosis DosS. J Biol Chem 291:16100-11
Matsumura, Hirotoshi; Chakraborty, Saumen; Reed, Julian et al. (2016) Effect of Outer-Sphere Side Chain Substitutions on the Fate of the trans Iron-Nitrosyl Dimer in Heme/Nonheme Engineered Myoglobins (Fe(B)Mbs): Insights into the Mechanism of Denitrifying NO Reductases. Biochemistry 55:2091-9

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