Abstract

The principal objective of this project is to understand in detail the mechanisms whereby Zn/Cu, Fe, and Mn containing superoxide dismutases catalyze the reaction, 202-+2H+ is greater than H2O2+O2. Our procedures involve the application of a variety of physical techniques ranging from x-ray crystallography to obtain three-dimensional structures to spectroscopic measuements to characterize the electronic structure of these metalloproteins. Kinetic mechanisms are being explored using a stopped-flow spectrophotometer specially designed for the study of superoxide in aqueous media. Simple recombinant DNA technqiues are being applied to the study of enzyme mechanisms and physiological function of these proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035189-04
Application #
3287510
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1984-12-01
Project End
1989-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Los Alamos National Lab
Department
Type
Organized Research Units
DUNS #
City
Los Alamos
State
NM
Country
United States
Zip Code
87545

  Publications

Tierney, D L; Fee, J A; Ludwig, M L et al. (1995) X-ray absorption spectroscopy of the iron site in Escherichia coli Fe(III) superoxide dismutase. Biochemistry 34:1661-8
Brown, O R; Smyk-Randall, E; Draczynska-Lusiak, B et al. (1995) Dihydroxy-acid dehydratase, a [4Fe-4S] cluster-containing enzyme in Escherichia coli: effects of intracellular superoxide dismutase on its inactivation by oxidant stress. Arch Biochem Biophys 319:10-22
Fee, J A (1991) Regulation of sod genes in Escherichia coli: relevance to superoxide dismutase function. Mol Microbiol 5:2599-610
Stallings, W C; Metzger, A L; Pattridge, K A et al. (1991) Structure-function relationships in iron and manganese superoxide dismutases. Free Radic Res Commun 12-13 Pt 1:259-68
Ludwig, M L; Metzger, A L; Pattridge, K A et al. (1991) Manganese superoxide dismutase from Thermus thermophilus. A structural model refined at 1.8 A resolution. J Mol Biol 219:335-58
Niederhoffer, E C; Naranjo, C M; Bradley, K L et al. (1990) Control of Escherichia coli superoxide dismutase (sodA and sodB) genes by the ferric uptake regulation (fur) locus. J Bacteriol 172:1930-8
Niederhoffer, E C; Fee, J A (1990) Novel effect of aromatic compounds on the iron-dependent expression of the Escherichia coli K12 manganese superoxide dismutase (sodA) gene. Biol Met 3:237-41
Carlioz, A; Ludwig, M L; Stallings, W C et al. (1988) Iron superoxide dismutase. Nucleotide sequence of the gene from Escherichia coli K12 and correlations with crystal structures. J Biol Chem 263:1555-62
Fee, J A; Bull, C (1986) Steady-state kinetic studies of superoxide dismutases. Saturative behavior of the copper- and zinc-containing protein. J Biol Chem 261:13000-5