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Catalytic Activity and Proton Translocation of Reconstituted Respiratory Complex I Monitored by Surface-Enhanced Infrared Absorption Spectroscopy
Citation key ISI:000433403900003
Author Gutierrez-Sanz, Oscar and Forbig, Enrico and Batista, Ana P. and Pereira, Manuela M. and Salewski, Johannes and Mroginski, Maria A. and Goetz, Robert and De Lacey, Antonio L. and Kozuch, Jacek and Zebger, Ingo
Pages 5703-5711
Year 2018
ISSN 0743-7463
DOI 10.1021/acs.langmuir.7b04057
Volume 34
Number 20
Month MAY 22
Abstract Respiratory complex I (CpI) is a key player in the way organisms obtain energy, being an energy transducer, which couples nicotinamide adenine dinucleotide (NADH)/quinone oxidoreduction with proton translocation by a mechanism that remains elusive so far. In this work, we monitored the function of CpI in a biomimetic, supported lipid membrane system assembled on a 4-aminothiophenol (4-ATP) self-assembled monolayer by surface-enhanced infrared absorption spectroscopy. 4-ATP serves not only as a linker molecule to a nanostructured gold surface but also as pH sensor, as indicated by concomitant density functional theory calculations. In this way, we were able to monitor NADH/quinone oxidoreduction-induced transmembrane proton translocation via the protonation state of 4-ATP, depending on the net onentation of CpI molecules induced by two complementary approaches. An associated change of the amide I/amide II band intensity ratio indicates conformational modifications upon catalysis which may involve movements of transmembrane helices or other secondary structural elements, as suggested in the literature.
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