This proposal describes how specific enzymes control the transfer of electrons and activation of molecular oxygen, while minimizing oxidative damage. This is central to cell development, health and survival. This project includes studies of enzyme reaction mechanisms, protein structure-function relationships, protein- protein interactions, protein post-translational modification, and mechanisms of long range biological electron transfer. Kinetic, biochemical, spectroscopic and structural studies together with site-directed mutagenesis will be used in these studies. This proposal focuses on the mechanism of biosynthesis ofthe protein-derived cofactor, tryptophan tryptophylquinone (TTQ), and the structure and function of a novel di-heme enzyme MauG which catalyzes the oxygenation and cross-linking of specific tryptophan residues during TTQ biogenesis in methylamine dehydrogenase (MADH). The substrate for MauG is a 119-kDa precursor protein of MADH with mono-hydroxylated Trp57 and no cross-link. MauG catalyzes the 6-electron oxidation ofthe substrate that results in the second oxygenation of Trp57, cross-linking of Trp57 and Trp108, and oxidation ofthe quinol product ofthe first two reactions to form oxidized TTQ. These studies will describe a new biological mechanism for oxygen activation and factors that make specific amino acid residues in proteins susceptible to oxidative modification.The results will provide insight for development of strategies to introduce novel catalytic sites into proteins and manipulate the functions of enzyme-bound hemes, as well as provide clues as to how one might mitigate naturally occurring oxidative damage to proteins. Ongoing mechanistic studies of biological electron transfer (ET) in the MADH-amicyanin-cytochrome c-551 i protein complex will be extended and ET studies will be initiated with MauG. Defining mechanisms of long range ET reactions will enhance our understanding ofthe fundamental processes of respiration and intermediary metabolism at the molecular level. A fundamental understanding ofthe mechanisms of control of biological ET reactions will provide insight into how defective protein ET leads to production of reactive oxygen species and free radicals both of which are associated with many disease states, oxidative stress and aging.

Public Health Relevance

Reactive oxygen species and free radicals produced as by-products of biological electron transfer and oxygen metabolism damage cell components that cause many disease states, oxidative stress and aging. However, free radicals and reactive oxygen species are also used productively in biosynthetic processes. These studies describe how specific enzymes control the transfer of electrons and activate oxygen, while m i n i m i z i n g o x i d a t i v e d a m a g e : . '. ^

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Method to Extend Research in Time (MERIT) Award (R37)
Project #
Application #
Study Section
No Study Section (in-house review) (NSS)
Program Officer
Barski, Oleg
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Central Florida
United States
Zip Code
Andreo-Vidal, Andres; Mamounis, Kyle J; Sehanobish, Esha et al. (2018) Structure and Enzymatic Properties of an Unusual Cysteine Tryptophylquinone-Dependent Glycine Oxidase from Pseudoalteromonas luteoviolacea. Biochemistry 57:1155-1165
Yukl, Erik T; Davidson, Victor L (2018) Diversity of structures, catalytic mechanisms and processes of cofactor biosynthesis of tryptophylquinone-bearing enzymes. Arch Biochem Biophys 654:40-46
Davidson, Victor L (2018) Protein-Derived Cofactors Revisited: Empowering Amino Acid Residues with New Functions. Biochemistry 57:3115-3125
Feng, Manliang; Ma, Zhongxin; Crudup, Breland F et al. (2017) Properties of the high-spin heme of MauG are altered by binding of preMADH at the protein surface 40 Å away. FEBS Lett 591:1566-1572
Ma, Zhongxin; Davidson, Victor L (2017) Ascorbate protects the diheme enzyme, MauG, against self-inflicted oxidative damage by an unusual antioxidant mechanism. Biochem J 474:2563-2572
Williamson, Heather R; Sehanobish, Esha; Shiller, Alan M et al. (2017) Roles of Copper and a Conserved Aspartic Acid in the Autocatalytic Hydroxylation of a Specific Tryptophan Residue during Cysteine Tryptophylquinone Biogenesis. Biochemistry 56:997-1004
Ma, Zhongxin; Williamson, Heather R; Davidson, Victor L (2016) A Suicide Mutation Affecting Proton Transfers to High-Valent Hemes Causes Inactivation of MauG during Catalysis. Biochemistry 55:5738-5745
Sehanobish, Esha; Campillo-Brocal, Jonatan C; Williamson, Heather R et al. (2016) Interaction of GoxA with Its Modifying Enzyme and Its Subunit Assembly Are Dependent on the Extent of Cysteine Tryptophylquinone Biosynthesis. Biochemistry 55:2305-8
Dow, Brian A; Davidson, Victor L (2016) Converting the bis-FeIV state of the diheme enzyme MauG to Compound I decreases the reorganization energy for electron transfer. Biochem J 473:67-72
Sehanobish, Esha; Williamson, Heather R; Davidson, Victor L (2016) Roles of Conserved Residues of the Glycine Oxidase GoxA in Controlling Activity, Cooperativity, Subunit Composition, and Cysteine Tryptophylquinone Biosynthesis. J Biol Chem 291:23199-23207

Showing the most recent 10 out of 56 publications