Our goal is to understand the fundamental electronic structure of paramagnetic metal and free radical proteins, to understand the interaction between naturally occuring paramagnetic centers in proteins, and to probe the dynamics of paramagnetic center motion. The outcome of the proposed work will be a more complete understanding of how the paramagnetic centers perform in their biological roles. The specific goals of the proposed research will be: 1. To study Cytochrome P-450 by electron nuclear double resonance (ENDOR) to determine its heme ligand environment. 2. To study by ENDOR the electronic structure at porphyrin-linked nitrogens and protons in ferric hemoglobins, coboglobins, and nitrosyl hemoglobins in order to detect effects of conformational change. 3. To probe by ENDOR the environs of the EPR-detectable """"""""copper"""""""" and heme in cytochrome c oxidase and to discover the nature of the ubisemiquinone radical in a newly found radical-ubiquinone protein of the electron transport chain. 4. To probe with pulsed EPR and saturation transfer EPR for evidence of inter-metal interaction and inter-metal distances in cytochrome c oxidase. 5. To look for the effect of slow molecular motions of O2 bound to coboglobins and of the ubiquinone in its protein environment. Pulsed EPR and saturation transfer EPR will be used. To perform these experiments we presently have a working ENDOR spectrometer and modicications for saturation transfer EPR. A pulsed saturation recovery apparatus is now being built to monitor electron spin-lattice relaxation, and a magnetic field pulsing device to operate in conjunction with it will be built as a part of the proposed work.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035103-15
Application #
3287210
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1984-12-01
Project End
1989-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
15
Fiscal Year
1989
Total Cost
Indirect Cost
Name
State University of New York at Albany
Department
Type
Schools of Arts and Sciences
DUNS #
City
Albany
State
NY
Country
United States
Zip Code
12222
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Barquera, Blanca; Morgan, Joel E; Lukoyanov, Dmitriy et al. (2003) X- and W-band EPR and Q-band ENDOR studies of the flavin radical in the Na+ -translocating NADH:quinone oxidoreductase from Vibrio cholerae. J Am Chem Soc 125:265-75
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DeWeerd, K; Grigoryants, V; Sun, Y et al. (2001) EPR-detected folding kinetics of externally located cysteine-directed spin-labeled mutants of iso-1-cytochrome c. Biochemistry 40:15846-55
Lukoyanov, D; Burger, R M; Scholes, C P (2001) ENDOR determination of the distance between bleomycin-bound iron and 19F of 2'-fluorocytidine in a DNA target sequence. J Am Chem Soc 123:12742-3
Veselov, A V; Osborne, J P; Gennis, R B et al. (2000) Q-band ENDOR (electron nuclear double resonance) of the high-affinity ubisemiquinone center in cytochrome bo3 from Escherichia coli. Biochemistry 39:3169-75
Sienkiewicz, A; da Costa Ferreira, A M; Danner, B et al. (1999) Dielectric resonator-based flow and stopped-flow EPR with rapid field scanning: A methodology for increasing kinetic information. J Magn Reson 136:137-42
Olesen, K; Veselov, A; Zhao, Y et al. (1998) Spectroscopic, kinetic, and electrochemical characterization of heterologously expressed wild-type and mutant forms of copper-containing nitrite reductase from Rhodobacter sphaeroides 2.4.3. Biochemistry 37:6086-94

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