Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase
| Autor: | Kevin M. Lewis, ChulHee Kang, Luying Xun, BuHyun Youn, Se-Young Jun |
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| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular 0301 basic medicine FMN Reductase Flavin Mononucleotide Stereochemistry Flavin group Microbiology Catalysis Article Cofactor Mixed Function Oxygenases Structure-Activity Relationship 03 medical and health sciences Oxidoreductase Catalytic Domain Flavins FMN reductase Flavin reductase NADH NADPH Oxidoreductases Amino Acid Sequence Enzyme kinetics Molecular Biology Edetic Acid chemistry.chemical_classification Oxidoreductases Acting on CH-NH Group Donors 030102 biochemistry & molecular biology biology Active site Phyllobacteriaceae Monooxygenase Hydroquinones Molecular Docking Simulation 030104 developmental biology chemistry Biochemistry biology.protein Oxidoreductases |
| Zdroj: | Molecular Microbiology. 100:989-1003 |
| ISSN: | 1365-2958 0950-382X |
| DOI: | 10.1111/mmi.13363 |
| Popis: | Ethylenediaminetetraacetate (EDTA) is currently the most abundant organic pollutant due to its recalcitrance and extensive use. Only a few bacteria can degrade it, using EDTA monooxygenase (EmoA) to initiate the degradation. EmoA is an FMNH2 -dependent monooxygenase that requires an NADH:FMN oxidoreductase (EmoB) to provide FMNH2 as a cosubstrate. Although EmoA has been identified from Chelativorans (ex. Mesorhizobium) sp. BNC1, its catalytic mechanism is unknown. Crystal structures of EmoA revealed a domain-like insertion into a TIM-barrel, which might serve as a flexible lid for the active site. Docking of MgEDTA(2-) into EmoA identified an intricate hydrogen bond network connected to Tyr(71) , which should potentially lower its pKa. Tyr(71) , along with nearby Glu(70) and a peroxy flavin, facilitates a keto-enol transition of the leaving acetyl group of EDTA. Further, for the first time, the physical interaction between EmoA and EmoB was observed by ITC, molecular docking and enzyme kinetic assay, which enhanced both EmoA and EmoB activities probably through coupled channelling of FMNH2 . |
| Databáze: | OpenAIRE |
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