The long range objectives of the research proposed in this application are to understand the catalytic mechanisms of several enzymes by using advanced electron paramagnetic resonance (EPR) spectroscopic methods to probe the detailed electronic structure of their catalytic sites and to gain an understanding of how these systems control chemical reactivity. The enzymes to be studied are copper-containing amine oxidases, methylamine dehydrogenases, methylmalonyl Coenzyme A Mutase, and carbonmonoxide dehydrogenase. All of these proteins catalyze reactions that involve paramagnetic intermediate states at some point during their cycle. This work will involve further development and use of pulsed and double resonance EPR experiments to measure magnetic and electronic interactions between the unpaired electron spin of the transient paramagnetic species and the magnetic nuclei that comprise their environment. Such hyperfine couplings provide a direct measurement of the highest occupied molecular orbital of the paramagnetic center and thus yield information on the valence electron distribution that is involved in catalysis. Such information is difficult or impossible to obtain by other spectroscopic methods and by X-ray crystallography. Such information will be correlated with the results of kinetic and other structural measurements. In particular the results will be related to electron transferring oxidation/reduction processes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM054065-03
Application #
2701732
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1996-05-01
Project End
1999-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
3
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Michigan State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Proshlyakov, Denis A; McCracken, John; Hausinger, Robert P (2017) Spectroscopic analyses of 2-oxoglutarate-dependent oxygenases: TauD as a case study. J Biol Inorg Chem 22:367-379
Cappillino, Patrick J; Miecznikowski, John R; Tyler, Laurie A et al. (2012) Studies of iron(II) and iron(III) complexes with fac-N2O, cis-N2O2 and N2O3 donor ligands: models for the 2-His 1-carboxylate motif of non-heme iron monooxygenases. Dalton Trans 41:5662-77
Sharpe, Martyn A; Krzyaniak, Matthew D; Xu, Shujuan et al. (2009) EPR evidence of cyanide binding to the Mn(Mg) center of cytochrome c oxidase: support for Cu(A)-Mg involvement in proton pumping. Biochemistry 48:328-35
Mills, Denise A; Xu, Shujuan; Geren, Lois et al. (2008) Proton-dependent electron transfer from CuA to heme a and altered EPR spectra in mutants close to heme a of cytochrome oxidase. Biochemistry 47:11499-509
McCracken, John; Vassiliev, Ilya R; Yang, En-Che et al. (2007) ESEEM studies of peptide nitrogen hyperfine coupling in tyrosyl radicals and model peptides. J Phys Chem B 111:6586-92
Velayutham, Murugesan; Muthukumaran, Rajendra B; Sostaric, Joe Z et al. (2007) Interactions of the major metabolite of the cancer chemopreventive drug oltipraz with cytochrome c: a novel pathway for cancer chemoprevention. Free Radic Biol Med 43:1076-85
Muthukumaran, Rajendra Bose; Grzyska, Piotr K; Hausinger, Robert P et al. (2007) Probing the iron-substrate orientation for taurine/alpha-ketoglutarate dioxygenase using deuterium electron spin echo envelope modulation spectroscopy. Biochemistry 46:5951-9
Schmidt, Bryan; Hillier, Warwick; McCracken, John et al. (2004) The use of stable isotopes and spectroscopy to investigate the energy transducing function of cytochrome c oxidase. Biochim Biophys Acta 1655:248-55
Henshaw, Timothy F; Feig, Michael; Hausinger, Robert P (2004) Aberrant activity of the DNA repair enzyme AlkB. J Inorg Biochem 98:856-61
Ryle, Matthew J; Liu, Aimin; Muthukumaran, Rajendra Bose et al. (2003) O2- and alpha-ketoglutarate-dependent tyrosyl radical formation in TauD, an alpha-keto acid-dependent non-heme iron dioxygenase. Biochemistry 42:1854-62

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