Abstract

Proposed research has a long-term goal to unravel the molecular mechanisms by which mutations of Sod1 gene encoding Cu, Zn superoxide dismutase (SOD1) produce selective motor neuron degeneration in amyotrophic lateral sclerosis (ALS) or Lou Gehrig's disease. The central hypothesis of this proposal is that bicarbonate anion enhances the peroxidative potential of SOD1 and its mutant variants via generation of a highly potent carbonate anion radical (CO3.-).
Specific Aims i nclude demonstration of CO3.- radical generation using purified enzymes; studies on the role of carbonate radical in hydroxylation, oxidation and nitration reactions; characterization of bicarbonate effects on oxidation/nitration reaction catalyzed by spinal cord extracts of transgenic ALS mutant mice and wild type animals. Four major analytical techniques to be used include electron spin resonance spectroscopy (ESR) and spin trapping; HPLC; UV-visible spectroscopy and chemiluminescence.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS040494-03
Application #
6540331
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (01))
Program Officer
Sheehy, Paul A
Project Start
2000-07-01
Project End
2004-05-31
Budget Start
2002-06-01
Budget End
2003-05-31
Support Year
3
Fiscal Year
2002
Total Cost
$167,668
Indirect Cost

  Institution

Name
Medical College of Wisconsin
Department
Biophysics
Type
Schools of Medicine
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226

  Publications

Chandran, Karunakaran; McCracken, John; Peterson, Francis C et al. (2010) Oxidation of histidine residues in copper-zinc superoxide dismutase by bicarbonate-stimulated peroxidase and thiol oxidase activities: pulse EPR and NMR studies. Biochemistry 49:10616-22
Zielonka, Jacek; Kalyanaraman, B (2010) Hydroethidine- and MitoSOX-derived red fluorescence is not a reliable indicator of intracellular superoxide formation: another inconvenient truth. Free Radic Biol Med 48:983-1001
Zielonka, Jacek; Hardy, Micael; Kalyanaraman, B (2009) HPLC study of oxidation products of hydroethidine in chemical and biological systems: ramifications in superoxide measurements. Free Radic Biol Med 46:329-38
Karunakaran, Chandran; Zhang, Hao; Joseph, Joy et al. (2005) Thiol oxidase activity of copper, zinc superoxide dismutase stimulates bicarbonate-dependent peroxidase activity via formation of a carbonate radical. Chem Res Toxicol 18:494-500
Pehar, Mariana; Cassina, Patricia; Vargas, Marcelo R et al. (2004) Astrocytic production of nerve growth factor in motor neuron apoptosis: implications for amyotrophic lateral sclerosis. J Neurochem 89:464-73
Zhang, Hao; Andrekopoulos, Christopher; Joseph, Joy et al. (2004) The carbonate radical anion-induced covalent aggregation of human copper, zinc superoxide dismutase, and alpha-synuclein: intermediacy of tryptophan- and tyrosine-derived oxidation products. Free Radic Biol Med 36:1355-65
Karunakaran, Chandran; Zhang, Hao; Crow, John P et al. (2004) Direct probing of copper active site and free radical formed during bicarbonate-dependent peroxidase activity of bovine and human copper, zinc-superoxide dismutases. Low-temperature electron paramagnetic resonance and electron nuclear double resonance stu J Biol Chem 279:32534-40
Zhang, Hao; Joseph, Joy; Crow, John et al. (2004) Mass spectral evidence for carbonate-anion-radical-induced posttranslational modification of tryptophan to kynurenine in human Cu, Zn superoxide dismutase. Free Radic Biol Med 37:2018-26
Vargas, Marcelo R; Pehar, Mariana; Cassina, Patricia et al. (2004) Stimulation of nerve growth factor expression in astrocytes by peroxynitrite. In Vivo 18:269-74
Andrekopoulos, Christopher; Zhang, Hao; Joseph, Joy et al. (2004) Bicarbonate enhances alpha-synuclein oligomerization and nitration: intermediacy of carbonate radical anion and nitrogen dioxide radical. Biochem J 378:435-47

Showing the most recent 10 out of 15 publications