Abstract

This program is devoted to the characterization of metalloenzyme active sites through the use of multinuclear, multifrequency (9; 35; 95 GHz) CW and pulsed electron-nuclear double resonance (ENDOR) and electron spin-echo envelope modulation (ESEEM) spectroscopies. The core of our research will be the study of oxygen-activating, Diron centers of, Hemes of, Methane Monoxygenase Nitric Oxide Synthase Ribonucleotide Reductase Cytochromes P450 This effort will emphasize the characterization of key catalytic intermediates. The uses of ENDOR/ESEEM will be expanded to the investigation of diamagnetic enzyme states, by making them paramagnetic through low-temperature reduction. Advances in resolution and sensitivity with our high-frequency pulsed ENDOR spectrometers will extend the reach of ENDOR beyond the primary metal ligands; substrate recognition will be analyzed through the determination of precise metrical structures for enzyme-bound substrates/inhibitors. These studies are linked by a desire to understand (i) the processes by which iron centers react with 02 to generate reactive high-valent [Fen- Om] species, (ii) the nature of these reactive species, and (iii) the means by which enzymes control their reaction with substrate.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL013531-31
Application #
6627504
Study Section
Metallobiochemistry Study Section (BMT)
Program Officer
Peterson, Charles M
Project Start
1979-01-01
Project End
2003-12-31
Budget Start
2003-01-01
Budget End
2003-12-31
Support Year
31
Fiscal Year
2003
Total Cost
$423,742
Indirect Cost

  Institution

Name
Northwestern University at Chicago
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
160079455
City
Evanston
State
IL
Country
United States
Zip Code
60201

  Publications

Davydov, Roman; Im, Sangchoul; Shanmugam, Muralidharan et al. (2016) Role of the Proximal Cysteine Hydrogen Bonding Interaction in Cytochrome P450 2B4 Studied by Cryoreduction, Electron Paramagnetic Resonance, and Electron-Nuclear Double Resonance Spectroscopy. Biochemistry 55:869-83
Shen, Jingmei; Kung, Mayfair C; Shen, Zhongliang et al. (2014) Generating and stabilizing Co(I) in a nanocage environment. J Am Chem Soc 136:5185-8
Davydov, Roman; Labby, Kristin Jansen; Chobot, Sarah E et al. (2014) Enzymatic and cryoreduction EPR studies of the hydroxylation of methylated N(?)-hydroxy-L-arginine analogues by nitric oxide synthase from Geobacillus stearothermophilus. Biochemistry 53:6511-9
Ortony, Julia H; Newcomb, Christina J; Matson, John B et al. (2014) Internal dynamics of a supramolecular nanofibre. Nat Mater 13:812-6
Hoffman, Brian M; Lukoyanov, Dmitriy; Yang, Zhi-Yong et al. (2014) Mechanism of nitrogen fixation by nitrogenase: the next stage. Chem Rev 114:4041-62
Lukoyanov, Dmitriy; Yang, Zhi-Yong; Duval, Simon et al. (2014) A confirmation of the quench-cryoannealing relaxation protocol for identifying reduction states of freeze-trapped nitrogenase intermediates. Inorg Chem 53:3688-93
Hoffman, Brian M; Lukoyanov, Dmitriy; Dean, Dennis R et al. (2013) Nitrogenase: a draft mechanism. Acc Chem Res 46:587-95
Sharma, Ajay; Gaidamakova, Elena K; Matrosova, Vera Y et al. (2013) Responses of Mn2+ speciation in Deinococcus radiodurans and Escherichia coli to ?-radiation by advanced paramagnetic resonance methods. Proc Natl Acad Sci U S A 110:5945-50
Davydov, Roman; Dawson, John H; Perera, Roshan et al. (2013) The use of deuterated camphor as a substrate in (1)H ENDOR studies of hydroxylation by cryoreduced oxy P450cam provides new evidence of the involvement of compound I. Biochemistry 52:667-71
Yang, Zhi-Yong; Khadka, Nimesh; Lukoyanov, Dmitriy et al. (2013) On reversible H2 loss upon N2 binding to FeMo-cofactor of nitrogenase. Proc Natl Acad Sci U S A 110:16327-32

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