Over the past decade it has been shown that very high frequency (VHF) EPR offers great advantages in many types of experiments. However, to date we have seen no documented attempts to apply this technique to viable biological systems. The reasons for this are both technical and psychological: the sample volume at W-band (95 GHz) could be 1000-fold smaller than at conventional X-band, and microwave losses are somewhat higher. In the present work we demonstrate the prospects for VHF EPR in cellular studies. The VHF EPR has a better absolute point sensitivity, which is essentially needed in studies of small objects (like a single cell) or surfaces, including cellular membranes. The better g-value resolution offered by VHF EPR enables one to separate spectroscopically different kinds of nitroxide radicals within the same system, or radicals in different molecular environments (e.g. to distinguish more clearly between nitroxide molecules dissolved in lipids, membranes, and aqueous phase of the cell). Another possible application of VHF EPR spectroscopy and imaging is in cryobiology, where nitroxide spin probes could be used to follow redistribution processes in cells during freezing.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001811-13
Application #
6120643
Study Section
Project Start
1998-04-15
Project End
1999-11-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Type
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Hurth, Kyle M; Nilges, Mark J; Carlson, Kathryn E et al. (2004) Ligand-induced changes in estrogen receptor conformation as measured by site-directed spin labeling. Biochemistry 43:1891-907
Woodmansee, Anh N; Imlay, James A (2002) Reduced flavins promote oxidative DNA damage in non-respiring Escherichia coli by delivering electrons to intracellular free iron. J Biol Chem 277:34055-66
Denisov, Ilia G; Makris, Thomas M; Sligar, Stephen G (2002) Formation and decay of hydroperoxo-ferric heme complex in horseradish peroxidase studied by cryoradiolysis. J Biol Chem 277:42706-10
Atsarkin, V A; Demidov, V V; Vasneva, G A et al. (2001) Mechanism of oxygen response in carbon-based sensors. J Magn Reson 149:85-9
Mangels, M L; Harper, A C; Smirnov, A I et al. (2001) Investigating magnetically aligned phospholipid bilayers with EPR spectroscopy at 94 GHz. J Magn Reson 151:253-9
Breitzer, J G; Smirnov, A I; Szczepura, L F et al. (2001) Redox properties of C6S8(n-) and C3S5(n-) (n = 0, 1, 2): stable radicals and unusual structural properties for C-S-S-C bonds. Inorg Chem 40:1421-9
Denisov, I G; Hung, S C; Weiss, K E et al. (2001) Characterization of the oxygenated intermediate of the thermophilic cytochrome P450 CYP119. J Inorg Biochem 87:215-26
Kirkor, E S; Scheeline, A (2000) Nicotinamide adenine dinucleotide species in the horseradish peroxidase-oxidase oscillator. Eur J Biochem 267:5014-22
Rapoport, N; Smirnov, A I; Pitt, W G et al. (1999) Bioreduction of Tempone and spin-labeled gentamicin by gram-negative bacteria: kinetics and effect of ultrasound. Arch Biochem Biophys 362:233-41
Maringanti, S; Imlay, J A (1999) An intracellular iron chelator pleiotropically suppresses enzymatic and growth defects of superoxide dismutase-deficient Escherichia coli. J Bacteriol 181:3792-802

Showing the most recent 10 out of 16 publications