Substrate-Protein Interactions of Type II NADH: Quinone Oxidoreductase from Escherichia coli

Johannes Salewski, Ana P. Batista, Filipa V. Sena, Diego Millo, Ingo Zebger, Manuela Alexandra Pereira, Peter Hildebrandt

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2722-2734
Number of pages13
JournalBiochemistry
Volume55
Issue number19
DOIs
Publication statusPublished - 17 May 2016

Fingerprint

Dive into the research topics of 'Substrate-Protein Interactions of Type II NADH: Quinone Oxidoreductase from Escherichia coli'. Together they form a unique fingerprint.

Cite this