Monoamine oxidases A and B (MAO A and B) are outer mitochondrial membrane-bound enzymes that function in the degradation of neurotransmitters such as serotonin, dopamine, and norepinephrine and have been pharmacologically important in the treatment of various pathological disorders such as depression, addition, and Parkinson's Disease. Each enzyme contains a covalent 8alpha-S-cysteinylFAD as a functional coenzyme. This project seeks continued support to determine the structure of each enzyme by x-ray crystallography to investigate the detailed mechanisms of H+ abstraction in catalysis. To facilitate these studies, we have developed methods for high level expression of each enzyme in Pichia pastrois. The role of the covalent FAD in MAO B will be investigated by disrupting the site for covalent FAD attachment by mutagenesis, followed by expression and purification of the mutant enzyme and comparison of its kinetic properties with WT enzyme The role of the C-terminal hydrophobic domain in enzyme structure and function for MAO A and B will be investigated in preparing the truncated forms and comparison of the purified mutants with WT enzyme properties. Initial studies are proposed to investigat3e the substrate specificity, inhibitor sensitivity, and structural properties of an expressed amine oxidase from Mycobacterium tuberculosis. Results from these studies should provide insights into the structures and mechanisms of these flavoprotein amine oxidases and lead to the development of new, specific drugs for the treatment of neuro-disorders and possibly tuberculosis infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029433-21
Application #
6729030
Study Section
Physical Biochemistry Study Section (PB)
Program Officer
Preusch, Peter C
Project Start
1982-07-01
Project End
2006-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
21
Fiscal Year
2004
Total Cost
$355,438
Indirect Cost
Name
Emory University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Edmondson, Dale E (2014) Hydrogen peroxide produced by mitochondrial monoamine oxidase catalysis: biological implications. Curr Pharm Des 20:155-60
Martinoli, Christian; Dudek, Hanna M; Orru, Roberto et al. (2013) Beyond the Protein Matrix: Probing Cofactor Variants in a Baeyer-Villiger Oxygenation Reaction. ACS Catal 3:3058-3062
Orru, R; Aldeco, M; Edmondson, D E (2013) Do MAO A and MAO B utilize the same mechanism for the C-H bond cleavage step in catalysis? Evidence suggesting differing mechanisms. J Neural Transm (Vienna) 120:847-51
Binda, Claudia; Aldeco, Milagros; Mattevi, Andrea et al. (2011) Interactions of monoamine oxidases with the antiepileptic drug zonisamide: specificity of inhibition and structure of the human monoamine oxidase B complex. J Med Chem 54:909-12
Wang, Jin; Edmondson, Dale E (2011) ²H kinetic isotope effects and pH dependence of catalysis as mechanistic probes of rat monoamine oxidase A: comparisons with the human enzyme. Biochemistry 50:7710-7
Aldeco, Milagros; Arslan, Betul Kacar; Edmondson, Dale E (2011) Catalytic and inhibitor binding properties of zebrafish monoamine oxidase (zMAO): comparisons with human MAO A and MAO B. Comp Biochem Physiol B Biochem Mol Biol 159:78-83
Milczek, Erika M; Binda, Claudia; Rovida, Stefano et al. (2011) The 'gating' residues Ile199 and Tyr326 in human monoamine oxidase B function in substrate and inhibitor recognition. FEBS J 278:4860-9
Wang, Jin; Edmondson, Dale E (2011) Topological probes of monoamine oxidases A and B in rat liver mitochondria: inhibition by TEMPO-substituted pargyline analogues and inactivation by proteolysis. Biochemistry 50:2499-505
Binda, Claudia; Mattevi, Andrea; Edmondson, Dale E (2011) Structural properties of human monoamine oxidases A and B. Int Rev Neurobiol 100:1-11
Binda, Claudia; Aldeco, Milagros; Geldenhuys, Werner J et al. (2011) Molecular Insights into Human Monoamine Oxidase B Inhibition by the Glitazone Anti-Diabetes Drugs. ACS Med Chem Lett 3:39-42

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