Print Email Facebook Twitter Revealing the Membrane-Bound Catalytic Oxidation of NADH by the Drug Target Type-II NADH Dehydrogenase Title Revealing the Membrane-Bound Catalytic Oxidation of NADH by the Drug Target Type-II NADH Dehydrogenase Author Godoy Hernandez, A. (TU Delft BT/Biocatalysis) Tate, Daniel J. (University of Manchester) McMillan, D.G.G. (TU Delft BT/Biocatalysis; University of Tokyo) Date 2019 Abstract Type-II NADH:quinone oxidoreductases (NDH-2s) are an important element of microbial pathogen electron transport chains and an attractive drug target. Despite being widely studied, its mechanism and catalysis are still poorly understood in a hydrophobic membrane environment. A recent report for the Escherichia coli NDH-2 showed NADH oxidation in a solution-based assay but apparently showed the reverse reaction in electrochemical studies, calling into question the validity of the electrochemical approach. Here we report electrochemical catalysis in the well-studied NDH-2 from Caldalkalibacillus thermarum (CthNDH-2). In agreement with previous reports, we demonstrated CthNDH-2 NADH oxidation in a solution assay and electrochemical assays revealed a system artifact in the absence of quinone that was absent in a membrane system. However, in the presence of either immobilized quinone or mobile quinone in a membrane, NADH oxidation was observed as in solution-phase assays. This conclusively establishes surface-based electrochemistry as a viable approach for interrogating electron transfer chain drug targets. To reference this document use: http://resolver.tudelft.nl/uuid:4d1a36bd-3430-49e6-9569-7e61207c3d98 DOI https://doi.org/10.1021/acs.biochem.9b00752 ISSN 0006-2960 Source Biochemistry, 58 (42), 4272-4275 Part of collection Institutional Repository Document type journal article Rights © 2019 A. Godoy Hernandez, Daniel J. Tate, D.G.G. McMillan Files PDF acs.biochem.9b00752.pdf 1.48 MB Close viewer /islandora/object/uuid:4d1a36bd-3430-49e6-9569-7e61207c3d98/datastream/OBJ/view