Molybdenum is an essential trace element that is found in enzymes involved in purine metabolism, sulfur metabolism, and the assimilation of inorganic nitrogen. Exposure to excess molybdenum has been linked with gout, and molybdenum levels in the soil affect the levels of nitrate and nitrite and also interfere with normal copper metabolism. In humans, fatal simultaneous deficiencies in enzyme activity for xanthine oxidase and sulfite oxidase have been shown to result from an inborn deficiency of the """"""""molybdenum cofactor,"""""""" an essential constituent of both enzymes. This research is directed toward understanding this common molybdenum cofactor, which is thought to be a monomeric molybdenum center with ligating sulfur atoms. One objective of this research is to synthesize and to structurally, chemically and spectroscopically characterize molybdenum complexes which are models for the molybdenum centers of these enzymes. Special emphasis will be given to complexes containing a monomeric oxo-molybdenum center linked to an iron porphyrin center as a model for sulfite oxidase. The compounds and their reactions with substrates and inhibitors of molybdoenzymes win be studied by electron paramagnetic resonance spectroscopy (EPR), spectroelectrochemistry, electronic spectroscopy and x-ray crystallography. A second objective is to directly investigate the molybdenum center of the enzyme sulfite oxidase by studying electron transfer processes between the molybdenum and iron centers of the enzyme and by continuing to grow crystals of sulfite oxidase with the ultimate goal of determining the three dimensional structure of the enzyme.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM037773-19
Application #
2178972
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1986-06-01
Project End
1995-11-30
Budget Start
1993-12-01
Budget End
1994-11-30
Support Year
19
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Arizona
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Kappler, Ulrike; Enemark, John H (2015) Sulfite-oxidizing enzymes. J Biol Inorg Chem 20:253-64
Klein, Eric L; Belaidi, Abdel Ali; Raitsimring, Arnold M et al. (2014) Pulsed electron paramagnetic resonance spectroscopy of (33)S-labeled molybdenum cofactor in catalytically active bioengineered sulfite oxidase. Inorg Chem 53:961-71
Davis, Amanda C; Johnson-Winters, Kayunta; Arnold, Anna R et al. (2014) Kinetic results for mutations of conserved residues H304 and R309 of human sulfite oxidase point to mechanistic complexities. Metallomics 6:1664-70
Davis, Amanda C; Cornelison, Matthew J; Meyers, Kimberly T et al. (2013) Effects of mutating aromatic surface residues of the heme domain of human sulfite oxidase on its heme midpoint potential, intramolecular electron transfer, and steady-state kinetics. Dalton Trans 42:3043-9
Johnson-Winters, Kayunta; Davis, Amanda C; Arnold, Anna R et al. (2013) Probing the role of a conserved salt bridge in the intramolecular electron transfer kinetics of human sulfite oxidase. J Biol Inorg Chem 18:645-53
Klein, Eric L; Astashkin, Andrei V; Raitsimring, Arnold M et al. (2013) Applications of pulsed EPR spectroscopy to structural studies of sulfite oxidizing enzymes(). Coord Chem Rev 257:110-118
Klein, Eric L; Raitsimring, Arnold M; Astashkin, Andrei V et al. (2012) Identity of the exchangeable sulfur-containing ligand at the Mo(V) center of R160Q human sulfite oxidase. Inorg Chem 51:1408-18
Astashkin, Andrei V; Rajapakshe, Asha; Cornelison, Matthew J et al. (2012) Determination of the distance between the Mo(V) and Fe(III) heme centers of wild type human sulfite oxidase by pulsed EPR spectroscopy. J Phys Chem B 116:1942-50
Rajapakshe, Asha; Meyers, Kimberly T; Berry, Robert E et al. (2012) Intramolecular electron transfer in sulfite-oxidizing enzymes: probing the role of aromatic amino acids. J Biol Inorg Chem 17:345-52
Rajapakshe, Asha; Tollin, Gordon; Enemark, John H (2012) Kinetic and thermodynamic effects of mutations of human sulfite oxidase. Chem Biodivers 9:1621-34

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