| Zitatschlüssel |
doi:10.1021/acs.biochem.6b00070 |
| Autor |
Salewski, Johannes and Batista, Ana P. and Sena, Filipa V. and Millo, Diego and Zebger, Ingo and Pereira, Manuela M. and Hildebrandt, Peter |
| Seiten |
2722-2734 |
| Jahr |
2016 |
| DOI |
10.1021/acs.biochem.6b00070 |
| Journal |
Biochemistry |
| Jahrgang |
55 |
| Nummer |
19 |
| Notiz |
PMID: 27109164 |
| Zusammenfassung |
Type II NADH:quinone oxidoreductases (NDH-2s) are membrane proteins involved in respiratory chains and responsible for the maintenance of NADH/NAD+ balance in cells. NDH-2s are the only enzymes with NADH dehydrogenase activity present in the respiratory chain of many pathogens, and thus, they were proposed as suitable targets for antimicrobial therapies. In addition, NDH-2s were also considered key players for the treatment of complex I-related neurodegenerative disorders. In this work, we explored substrate–protein interaction in NDH-2 from Escherichia coli (EcNDH-2) combining surface-enhanced infrared absorption spectroscopic studies with electrochemical experiments, fluorescence spectroscopy assays, and quantum chemical calculations. Because of the specific stabilization of substrate complexes of EcNDH-2 immobilized on electrodes, it was possible to demonstrate the presence of two distinct substrate binding sites for NADH and the quinone and to identify a bound semiprotonated quinol as a catalytic intermediate. |
