2D-SEIRA spectroscopy to highlight conformational changes of the cytochrome c oxidase induced by direct electron transfer

Christoph Nowak; Thamara Laredo; Jens Gebert; Jacek Lipkowski; Robert B. Gennis; Shelagh Ferguson-Miller; Wolfgang Knoll; Renate L. C. Naumann

doi:10.1039/C0MT00083C

2D-SEIRAspectroscopy to highlight conformational changes of the cytochrome coxidase induced by direct electron transfer†‡

Christoph Nowak,^a Thamara Laredo,^b Jens Gebert,^a Jacek Lipkowski,^c Robert B. Gennis,^d Shelagh Ferguson-Miller,^e Wolfgang Knoll^a and Renate L. C. Naumann*^a

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* Corresponding authors

^a Austrian Institute of Technology GmbH, AIT, Donau-City Str. 1, 1220 Vienna, Austria
E-mail: C.Nowak@ait.ac.at, Jens.Gebert.fl@ait.ac.at, Wolfgang.Knoll@ait.ac.at, Renate.Naumann@ait.ac.at

^b Lakehead University, Orillia, Canada
E-mail: tlaredo1@lakeheadu.ca

^c University of Guelph, Department of Chemistry and Biochemistry, Guelph, Canada
E-mail: jlipkows@uoguelph.ca

^d University of Illinois, Department of Biochemistry, 600 South Mathews Street, Urbana, USA
E-mail: r-gennis@illinois.edu

^e Michigan State University Biochemistry and Molecular Biology, East Lansing, MI, USA
E-mail: fergus20@mail.msu.edu

Abstract

Potentiometric titrations of the cytochrome c oxidase (CcO) immobilized in a biomimetic membrane system were followed by two-dimensional surface-enhanced IR absorption spectroscopy (2D SEIRAS) in the ATR-mode. Direct electron transfer was employed to vary the redox state of the enzyme. The CcO was shown to undergo a conformational transition from a non-activated to an activated state after it was allowed to turnover in the presence of oxygen. Differences between the non-activated and activated state were revealed by 2D SEIRA spectra recorded as a function of potential. The activated state was characterized by a higher number of correlated transitions as well as a higher number of amino acids associated with electron transfer.

Metallomics

2D-SEIRAspectroscopy to highlight conformational changes of the cytochrome coxidase induced by direct electron transfer†‡

Abstract

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