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SHR Neuro Cancer Cardio Lipid Metab Microb
Sena, FV; Batista, AP; Catarino, T; Brito, JA; Archer, M; Viertler, M; Madl, T; Cabrita, EJ; Pereira, MM.
Type-II NADH:quinone oxidoreductase from Staphylococcus aureus has two distinct binding sites and is rate limited by quinone reduction.
Mol Microbiol. 2015; 98(2):272-288
Doi: 10.1111/mmi.13120
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- Co-authors Med Uni Graz
- Madl Tobias
- Viertler Martin
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- Abstract:
- A prerequisite for any rational drug design strategy is understanding the mode of protein-ligand interaction. This motivated us to explore protein-substrate interaction in Type-II NADH:quinone oxidoreductase (NDH-2) from Staphylococcus aureus, a worldwide problem in clinical medicine due to its multiple drug resistant forms. NDHs-2 are involved in respiratory chains and recognized as suitable targets for novel antimicrobial therapies, as these are the only enzymes with NADH:quinone oxidoreductase activity expressed in many pathogenic organisms. We obtained crystal and solution structures of NDH-2 from S. aureus, showing that it is a dimer in solution. We report fast kinetic analyses of the protein and detected a charge-transfer complex formed between NAD(+) and the reduced flavin, which is dissociated by the quinone. We observed that the quinone reduction is the rate limiting step and also the only half-reaction affected by the presence of HQNO, an inhibitor. We analyzed protein-substrate interactions by fluorescence and STD-NMR spectroscopies, which indicate that NADH and the quinone bind to different sites. In summary, our combined results show the presence of distinct binding sites for the two substrates, identified quinone reduction as the rate limiting step and indicate the establishment of a NAD(+)-protein complex, which is released by the quinone. © 2015 John Wiley & Sons Ltd.
- Find related publications in this database (using NLM MeSH Indexing)
- Bacterial Proteins - chemistry
- Bacterial Proteins - genetics
- Bacterial Proteins - metabolism
- Binding Sites -
- Crystallography, X-Ray -
- Drug Discovery -
- Electron Transport -
- Hydroxyquinolines - pharmacology
- Kinetics -
- Models, Molecular -
- Oxidation-Reduction -
- Protein Multimerization -
- Quinone Reductases - antagonists & inhibitors
- Quinone Reductases - chemistry
- Quinone Reductases - genetics
- Quinone Reductases - metabolism
- Quinones - metabolism
- Staphylococcus aureus - enzymology
- Staphylococcus aureus - metabolism