The PI proposes to investigate the 3D structure, active site architecture, catalytic mechanism and mechanism of activation of soluble methane monooxygenase from the Type II methanotroph Methylosinus trichosporum OB3b. This enzyme catalyzes the first step in the oxidation of methane to CO2 by methanogenic bacteria. In this way, the atmospheric egress of nearly all the enormous quantity of biogenic methane (a potent greenhouse gas) generated by anaerobic bacteria is prevented. MMO also adventitiously catalyzes the oxidation of many other hydrocarbons leading to applications in synthesis and biodegradation. Work to date suggests that the reaction is catalyzed by a cofactor not found in other oxygenases, implying a new strategy for oxygen activation. The proposed studies build on the very significant progress that has been made in the previous funding period, and focus on: elucidation of the physical and electronic structure of dinuclear iron cluster of the active site as well as the mechanism of oxygen activation catalyzed by this center; spectroscopic and kinetic characterization of the several reaction intermediates that have been identified in the catalytic sequence; elucidation of the protein-protein interactions between the three polypeptides of the enzyme that modulate its catalytic activity.
| Komor, Anna J; Jasniewski, Andrew J; Que, Lawrence et al. (2018) Diiron monooxygenases in natural product biosynthesis. Nat Prod Rep 35:646-659 |
| Oloo, Williamson N; Banerjee, Rahul; Lipscomb, John D et al. (2017) Equilibrating (L)FeIII-OOAc and (L)FeV(O) Species in Hydrocarbon Oxidations by Bio-Inspired Nonheme Iron Catalysts Using H2O2 and AcOH. J Am Chem Soc 139:17313-17326 |
| Castillo, Rebeca G; Banerjee, Rahul; Allpress, Caleb J et al. (2017) High-Energy-Resolution Fluorescence-Detected X-ray Absorption of the Q Intermediate of Soluble Methane Monooxygenase. J Am Chem Soc 139:18024-18033 |
| Banerjee, Rahul; Proshlyakov, Yegor; Lipscomb, John D et al. (2015) Structure of the key species in the enzymatic oxidation of methane to methanol. Nature 518:431-4 |
| Lipscomb, John D (2014) Life in a sea of oxygen. J Biol Chem 289:15141-53 |
| Banerjee, Rahul; Meier, Katlyn K; Munck, Eckard et al. (2013) Intermediate P* from soluble methane monooxygenase contains a diferrous cluster. Biochemistry 52:4331-42 |
| Makris, Thomas M; Knoot, Cory J; Wilmot, Carrie M et al. (2013) Structure of a dinuclear iron cluster-containing ?-hydroxylase active in antibiotic biosynthesis. Biochemistry 52:6662-71 |
| Vu, Van V; Makris, Thomas M; Lipscomb, John D et al. (2011) Active-site structure of a ýý-hydroxylase in antibiotic biosynthesis. J Am Chem Soc 133:6938-41 |
| Makris, Thomas M; Chakrabarti, Mrinmoy; Münck, Eckard et al. (2010) A family of diiron monooxygenases catalyzing amino acid beta-hydroxylation in antibiotic biosynthesis. Proc Natl Acad Sci U S A 107:15391-6 |
| Mitic, Natasa; Schwartz, Jennifer K; Brazeau, Brian J et al. (2008) CD and MCD studies of the effects of component B variant binding on the biferrous active site of methane monooxygenase. Biochemistry 47:8386-97 |
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