Blaza, James N orcid.org/0000-0001-5420-2116, Bridges, Hannah R, Aragão, David et al. (5 more authors) (2017) The mechanism of catalysis by type-II NADH:quinone oxidoreductases. Scientific Reports. 40165. ISSN 2045-2322
Abstract
Type II NADH:quinone oxidoreductase (NDH-2) is central to the respiratory chains of many organisms. It is not present in mammals so may be exploited as an antimicrobial drug target or used as a substitute for dysfunctional respiratory complex I in neuromuscular disorders. NDH-2 is a single-subunit monotopic membrane protein with just a flavin cofactor, yet no consensus exists on its mechanism. Here, we use steady-state and pre-steady-state kinetics combined with mutagenesis and structural studies to determine the mechanism of NDH-2 from Caldalkalibacillus thermarum. We show that the two substrate reactions occur independently, at different sites, and regardless of the occupancy of the partner site. We conclude that the reaction pathway is determined stochastically, by the substrate/product concentrations and dissociation constants, and can follow either a ping-pong or ternary mechanism. This mechanistic versatility provides a unified explanation for all extant data and a new foundation for the development of therapeutic strategies.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s) 2017 |
Keywords: | Bacillales,Binding Sites,Catalysis,Dinitrocresols/metabolism,Kinetics,Protein Binding,Quinone Reductases/chemistry,Reactive Oxygen Species/metabolism |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Depositing User: | Pure (York) |
Date Deposited: | 21 Nov 2018 13:30 |
Last Modified: | 07 Feb 2025 00:23 |
Published Version: | https://doi.org/10.1038/srep40165 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1038/srep40165 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:138995 |