Project A1

Electron-driven protonation dynamics in cytochrome c oxidase

Principal Investigators: Prof. Dr. Joachim Heberle (FU), Prof. Dr. Inez Weidinger (TU)

The A1 project aims at the comprehensive monitoring of electron-driven proton translocations in oxidases (cytochrome c and ubiquinol oxidases) by time-resolved infrared (IR) and resonance Raman spectroscopy. Novel techniques will be developed and applied to initiate and monitor protonation dynamics: Microfluidic flow-flash setup for triggering the reaction by a short laser pulse as well as electrochemical electron injection to solid-supported oxidase in combination with surface-enhanced IR and resonance Raman spectroscopies. By these means, the reaction sequence, the rate-limiting steps and the key residues in CcO will be identified.

Publications 2013 - 2018

Ataka, K., Stripp, S.T., and Heberle, J. (2013). Surface-enhanced infrared absorption spectroscopy (SEIRAS) to probe monolayers of membrane proteins. Biochimica et Biophysica Acta (BBA) - Biomembranes 1828, 2283-2293.

Kielb, P., Sezer, M., Katz, S., Lopez, F., Schulz, C., Gorton, L., Ludwig, R., Wollenberger, U., Zebger, I., and Weidinger, I.M. (2015). Spectroscopic Observation of Calcium-Induced Reorientation of Cellobiose Dehydrogenase Immobilized on Electrodes and its Effect on Electrocatalytic Activity. ChemPhysChem 16, 1960-1968.

Kielb, P., Utesch, T., Kozuch, J., Jeoung, J.-H., Dobbek, H., Mroginski, M.A., Hildebrandt, P., and Weidinger, I. (2017). Switchable Redox Chemistry of the Hexameric Tyrosine-Coordinated Heme Protein. Journal of Physical Chemistry B 121, 3955-3964.

Kottke, T., Lórenz-Fonfría V.A., and Heberle J. (2017). The Grateful Infrared – Sequential Protein Structural Changes Resolved by IR Difference Spectroscopy. Journal of Physical Chemistry B 121, 335-350. [Link to news section.]

Kozuch, J., von der Hocht, I., Hilbers, F., Michel, H., and Weidinger, I.M. (2013). Resonance Raman Characterization of the Ammonia-Generated Oxo Intermediate of Cytochrome c Oxidase from Paracoccus denitrificans. Biochemistry 52, 6197-6202.

Lorenz-Fonfria V.A., Saita M., Lazarova T., Schlesinger R., and Heberle J. (2017). pH-sensitive vibrational probe reveals a cytoplasmic protonated cluster in bacteriorhodopsin. Proceedings of the National Academy of Sciences of the United States of America 114, E10909-E10918

Ly, H.K., Wrzolek, P., Heidary, N., Gotz, R., Horch, M., Kozuch, J., Schwalbe, M., and Weidinger, I.M. (2015). 2nd coordination sphere controlled electron transfer of iron hangman complexes on electrodes probed by surface enhanced vibrational spectroscopy. Chem. Sci. 6, 6999-7007.

Mohrmann, H., Kube, I., Lórenz-Fonfría, V.A., Engelhard, M., and Heberle, J. (2016). Transient Conformational Changes of Sensory Rhodopsin II Investigated by Vibrational Stark Effect Probes. Journal of Physical Chemistry B 120, 4383-4387.

Schultz, B.-J., Mohrmann, H., Lórenz-Fonfría, V.A., and Heberle J. (2018). Protein dynamics observed by tunable mid-IR quantum cascade lasers across the time range from 10 ns to 1 s. Spectrochimica Acta A 188, 666-674.

Sezer, M., Kielb, P., Kuhlmann, U., Mohrmann, H., Schulz, C., Heinrich, D., Schlesinger, R., Heberle, J., and Weidinger, I.M. (2015). Surface Enhanced Resonance Raman Spectroscopy Reveals Potential Induced Redox and Conformational Changes of Cytochrome c Oxidase on Electrodes. Journal of Physical Chemistry B 119, 9586-9591.

Sezer, M., Woelke, A.-L., Knapp, E.W., Schlesinger, R., Mroginski, M.A., Weidinger, I.M. (2017). Redox induced protonation of heme propionates in cytochrome c oxidase: Insights from surface enhanced resonance Raman spectroscopy and QM/MM calculations. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1858, 103-108.

Süss, B., Ringleb, F., and Heberle, J. (2016). New ultrarapid-scanning interferometer for FT-IR spectroscopy with microsecond time-resolution. Review of Scientific Instruments 87, 063113.

Weidinger, I.M. (2015). Analysis of structure–function relationships in cytochrome c oxidase and its biomimetic analogs via resonance Raman and surface enhanced resonance Raman spectroscopies. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1847, 119-125.

Woelke, A.L., Galstyan, G., Galstyan, A., Meyer, T., Heberle, J., and Knapp, E.-W. (2013). Exploring the Possible Role of Glu286 in CcO by Electrostatic Energy Computations Combined with Molecular Dynamics. Journal of Physical Chemistry B 117, 12432-12441.