Hydrogen bonding network in channel- and other rhodopsins and phytochromes analyzed by site-directed mutagenesis and labeling

Principal Investigators: Dr. Ramona Schlesinger (FU)

We will comparatively study the protonation dynamics of two different channelrhodopsins, CaChR1 and GtACR1, the inverted proton pump NsXeR and the chloride pump NmHR in a combination of mutagenesis, time-resolved UV/Vis-, steady-state FTIR difference spectroscopy and structural methods. Local electric field changes will be observed via VSE probes inserted into the two pumps and into phytochromes. In the complex of HsSRI and HsHtrI, we will study the structural changes involved in signal transfer.

Publications

2021 - 2024

Ataka, K., Baumann, A., Chen, J.L., Redlich, A., Heberle, J. and Schlesinger, R. (2022). Monitoring the Progression of Cell-Free Expression of Microbial Rhodopsins by Surface Enhanced IR Spectroscopy: Resolving a Branch Point for Successful/Unsuccessful Folding. Front. Mol. Biosci., 9, 929285. doi: 10.3389/fmolb.2022.929285.

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.

Bühl, E., Resler, T., Lam, R., Asido, M., Bamberg, E., Schlesinger, R., Bamann, C., Heberle, J. and Wachtveitl, J. (2023). Assessing the Role of R120 in the Gating of CrChR2 by Time-Resolved Spectroscopy from Femtoseconds to Seconds. J Am Chem Soc 145, 40: 21832-21840. doi: 10.1021/jacs.3c05399.

Crea, F., Vorkas, A., Redlich, A., Cruz, R., Shi, C., Trauner, D., Lange, A., Schlesinger, R. and Heberle, J. (2022). Photoactivation of a Mechanosensitive Channel. Front. Mol. Biosci. doi: 10.3389/fmolb.2022.905306. 

La Greca, M., Chen, J.-L., Schubert, L., Kozuch, J., Berneiser, T., Terpitz, U., Heberle, J., and Schlesinger, R. (2022). The Photoreaction of the Proton-Pumping Rhodopsin 1 From the Maize Pathogenic Basidiomycete Ustilago maydis. Front. Mol. Biosci. doi: 10.3389/fmolb.2022.826990.

Schubert, L., Chen, J.-L., Fritz, T., Marxer, F., Langner, P., Hoffmann, K., Gamiz-Hernandez, A.P., Kaila, V.R.I., Schlesinger, R. and Heberle, J. (2023). Proton Release Reactions in the Inward H+ Pump NsXeR. J. Phys. Chem. B. doi: 10.1021/acs.jpcb.3c04100.

Walter, M., and Schlesinger, R. (2021). Nanodisc Reconstitution of Channelrhodopsins Heterologously Expressed in Pichia pastoris for Biophysical Investigations. In: Dempski R. (eds) Channelrhodopsin. Methods. Mol. Biol., 2191. Humana, New York, NY. doi: 10.1007/978-1-0716-0830-2_3.

Walter, M., Schubert, L., Heberle, J., Schlesinger, R. and Losi, A. (2023). Time-resolved photoacoustics of channelrhodopsins: early energetics and light-driven volume changes. Photochem. Photobiol. Sci. doi: 10.1007/s43630-022-00327-8.

Yaguchi, M., Jia, X., Schlesinger, R., Jiang, X., Ataka, K., and Heberle, J. (2022). Near-Infrared Activation of Sensory Rhodopsin II Mediated by NIR-to-Blue Upconversion Nanoparticles. Front. Mol. Biosci. doi: 10.3389/fmolb.2021.782688.

2017 - 2020

Ehrenberg, D., Krause, N., Saita, M., Bamann, C., Kar, R.K., Hoffmann, K., Heinrich, D., Schapiro, I., Heberle, J., and Schlesinger, R. (2019) Atomistic insight into the role of threonine 127 in the functional mechanism of channelrhodopsin-2. Appl. Sci., 9 (22), 4905; https://doi.org/10.3390/app9224905.

Harris N.J., Reading E., Ataka K., Grzegorzewski L., Charalambous K., Liu X., Schlesinger R., Heberle J., and Booth P.J. (2017). Structure formation during translocon-unassisted co-translational membrane protein folding. Scientific Reports 7: 8021; doi: 10.1038/s41598-017-08522-9

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

Kovácsová, G., Grunbein, M.L., Kloos, M., Barends, T.R.M., Schlesinger, R., Heberle, J., Kabsch, W., Shoeman, R.L., Doak, R.B., and Schlichting, I. (2017). Viscous hydrophilic injection matrices for serial crystallography. IUCrJ 4, 400-410; doi: 10.1107/S2052252517005140

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: 10.1038/s41598-020-60801-0

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; doi: 10.1073/pnas.1707993114

Nass Kovacs, G., Colletier, J.-P., Grünbein, M. L., Yang, Y., Stensitzki, T., Batyuk, A., Carbajo, S., Doak, R. B., Ehrenberg, D., Foucar, L., Gasper, R., Gorel, A., Hilpert, M., Kloos, M., Koglin, J. E., Reinstein, J., Roome, C. M., Schlesinger, R., Seaberg, M., Shoeman, R. L., Stricker, M., Boutet, S., Haacke, S., Heberle, J., Heyne, K., Domratcheva, T., Barends, T. R. M., and Schlichting, I.(2019). Three-dimensional view of ultrafast dynamics in photoexcited bacteriorhodopsin. Nature Communications 10, Article Number 3177. doi: 10.1038/s41467-019-10758-0

Pfitzner, E., Seki, H., Schlesinger, R., Ataka, K., and Heberle, J. (2018). Disc Antenna Enhanced Infrared Spectroscopy: From Self-Assembled Monolayers to Membrane Proteins. ACS Sens, 3 (5), pp 984–991, doi:10.1021/acssensors.8b00139.

Saita, M., Pranga-Sellnau, F., Resler, T., Schlesinger, R., Heberle, J., and Lorenz-Fonfria, V.A. (2018). Photoexcitation of the P4(480) State Induces a Secondary Photocycle That Potentially Desensitizes Channelrhodopsin-2. J. Am. Chem. Soc. 140, 9899-9903; doi: 10.1021/jacs.8b03931

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: 10.1016/j.bbabio.2016.10.009

Stensitzki, T., Adam, S., Schlesinger, R., Schapiro, I., and Heyne, K. (2020). Ultrafast Backbone Protonation in Channelrhodopsin-1 Captured by Polarization Resolved Fs Vis-pump-IR-Probe Spectroscopy and Computational Methods. Molecules 25, 848; doi: 10.3390/molecules25040848

Weber A. M., Kaiser, J., Ziegler, T., Pilsl, S.,, Renzl, C., Sixt, L., Pietruschka, G., Moniot, S., Kakoti, A., Juraschitz, M., Schrottke, S.,Lledo Bryant, L., Steegborn, C., Bittl, R., Mayer, G., and Möglich, A. (2019). A blue light receptor that mediates RNA binding and translational regulation. Nature Chemical Biology, doi: 10.1038/s41589-019-0346-y

2013 - 2016

Krause, N., Engelhard, C., Heberle, J., Schlesinger, R., and Bittl, R. (2013). Structural differences between the closed and open states of channelrhodopsin-2 as observed by EPR spectroscopy. FEBS Letters 587, 3309-3313.

Lórenz-Fonfría, V.A., Bamann, C., Resler, T., Schlesinger, R., Bamberg, E., and Heberle, J. (2015). Temporal evolution of helix hydration in a light-gated ion channel correlates with ion conductance. Proceedings of the National Academy of Sciences of the United States of America 112, E5796-E5804.

Lórenz-Fonfría, V.A., Muders, V., Schlesinger, R., and Heberle, J. (2014). Changes in the hydrogen-bonding strength of internal water molecules and cysteine residues in the conductive state of channelrhodopsin-1. Journal of Chemical Physics 141, 22D507.

Lorenz-Fonfria, V.A., Resler, T., Krause, N., Nack, M., Gossing, M., von Mollard, G.F., Bamann, C., Bamberg, E., Schlesinger, R., and Heberle, J. (2013). Transient protonation changes in channelrhodopsin-2 and their relevance to channel gating. Proceedings of the National Academy of Sciences of the United States of America 110, E1273-E1281.

Lórenz-Fonfría, V.A., Schultz, B.-J., Resler, T., Schlesinger, R., Bamann, C., Bamberg, E., and Heberle, J. (2015). Pre-Gating Conformational Changes in the ChETA Variant of Channelrhodopsin-2 Monitored by Nanosecond IR Spectroscopy. Journal of the American Chemical Society 137, 1850-1861.

Muders, V., Kerruth, S., Lorenz-Fonfria, V.A., Bamann, C., Heberle, J., and Schlesinger, R. (2014). Resonance Raman and FTIR spectroscopic characterization of the closed and open states of channelrhodopsin-1. FEBS Letters 588, 2301-2306.

Resler, T., Schultz, B.-J., Lorenz-Fonfria, V.A., Schlesinger, R., and Heberle, J. (2015). Kinetic and Vibrational Isotope Effects of Proton Transfer Reactions in Channelrhodopsin-2. Biophysical Journal 109, 287-297.

Schnedermann, C., Muders, V., Ehrenberg, D., Schlesinger, R., Kukura, P., and Heberle, J. (2016). Vibronic Dynamics of the Ultrafast all-trans to 13-cis Photoisomerization of Retinal in Channelrhodopsin-1. Journal of the American Chemical Society 138, 4757-4762.

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.

Stensitzki, T., Muders, V., Schlesinger, R., Heberle, J., and Heyne, K. (2015). The primary photoreaction of channelrhodopsin-1: Wavelength dependent photoreactions induced by ground-state heterogeneity. Frontiers in Molecular Biosciences 2, 41.

Stensitzki, T., Yang, Y., Muders, V., Schlesinger, R., Heberle, J., and Heyne, K. (2016). Femtosecond infrared spectroscopy of channelrhodopsin-1 chromophore isomerization. Structural Dynamics 3, 043208.

Volz, P., Krause, N., Balke, J., Schneider, C., Walter, M., Schneider, F., Schlesinger, R., and Alexiev, U. (2016). Light and pH-induced changes in structure and accessibility of transmembrane helix B and its immediate environment in Channelrhodopsin-2. The Journal of Biological Chemistry 291, 17382-17393.