Electron-driven protonation dynamics in cytochrome c oxidase

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

In this project, state-of-the-art vibrational spectroscopic methods are applied to obtain a molecular understanding of the interplay between redox chemistry, oxygen reduction and protonation dynamics in cytochrome c oxidase. By combining resonance Raman and IR spectroscopy with electrochemical techniques, we will monitor key functional groups in proton translocation such as Y288, R473 and heme propionates on the μs-s time scale and as a function of the applied potential.

Publications

2021-2024

Baserga, F., Dragelj, J., Kozuch, J., Mohrmann, H., Knapp, E.-W., Stripp, S. T., and Heberle, J. (2021). Quantification of local electric field changes at the active site of Cytochrome c Oxidase by Fourier transform infrared spectroelectrochemical titrations. Front. Chem. 9, 669452. doi: 10.3389/fchem.2021.669452.

Baserga, F., Storm, J., Schlesinger, R., Heberle, J. and Stripp, S.T. (2023). The catalytic reaction of cytochrome c oxidase probed by in situ gas titrations and FTIR difference spectroscopy. Biochim. Biophys. Acta, 1864, 4: 149000. doi: 10.1016/j.bbabio.2023.149000.

Baserga, F., Vorkas, A., Crea, F., Schubert, L., Chen, J.-L., Redlich, A., La Greca, M., Storm, J., Oldemeyer, S., Hoffmann, K., Schlesinger, R. and Heberle, J. (2022). Membrane Protein Activity Induces Specific Molecular Changes in Nanodiscs Monitored by FTIR Difference Spectroscopy. Front Mol Biosci, 9. doi: 10.3389/fmolb.2022.915328.

Göbel, M., Ramuglia, A.R., Zhartovska, E., Budhija, V., Ly, K.H., Mroginski, M.A., Schwalbe, M. and Weidinger, I.M. (2024). Protonation of Pendant Pyridine Substituents in an Iron Porphyrin Hangman Complex: Influence on Spectral Visibility and Electrocatalysis. J Phys Chem C, 128, 18: 7624-7634. doi: 10.1021/acs.jpcc.4c01357.

Kruse, F., Nguyen, A.D., Dragelj, J., Heberle, J., Hildebrandt, P., Mroginski, M.A., and Weidinger, I.M. (2021). A Resonance Raman Marker Band Characterizes the Slow and Fast Form of Cytochrome c Oxidase. J Am Chem Soc, 143, 2769-2776. doi: 10.1021/jacs.0c10767.

2017-2020

Hugentobler, K., Heinrich, D., Berg, J., Heberle, J., Brzezinski, P., Schlesinger, R., Block, S. (2020) Lipid Composition Affects the Efficiency in the Functional Reconstitution of the Cytochrome c Oxidase. Int. J. Mol. Sci. , 21(19), 6981; https://doi.org/10.3390/ijms21196981

Kielb, P., Horch, M., Wrzolek, P., Goetz, R., Ly, H.K., Kozuch, J., Schwalbe, M. and Weidinger, I.M. (2018) Hydrogen evolution by cobalt Hangman porphyrins under operating conditions studied by vibrational spectro-electrochemistry. Catal. Sci. Technol. 8, 1849-1857; doi.org/10.1039/C7CY02253K

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. doi.org/10.1021/acs.jpcb.7b01286

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.]

Kruse, F., Nguyen, A.D., Dragelj, J., Schlesinger, R., Heberle, J., Mroginski, M.A. and Weidinger, I.M. (2020). Characterisation of the Cyanate Inhibited State of Cytochrome c Oxidase. Sci. Rep. 10, 3863; doi.org/10.1038/s41598-020-60801-0

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; doi.org/10.1016/j.saa.2017.01.010

Senger, M., Eichmann, V., Laun, K., Duan, J., Wittkamp, F., Knör, G., Apfel, U.-P., Happe, T., Winkler, M., Heberle, J., Stripp, S. T. (2019). How [FeFe]-Hydrogenase Facilitates Bidirectional Proton Transfer. J. Am. Chem. Soc. 141 (43), 17394-17403; doi.org/10.1021/jacs.9b09225.

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. doi.org/10.1016/j.bbabio.2016.10.009

2013-2016

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.

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.

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.

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.

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.