Project B3

Video on time-resolved step-scan FT-IR in J. Vis. Exp. (June 2014)
Video by Lorenz-Fonfria and Heberle on Time-Resolved Step-Scan FT-IR in J. Vis. Exp. (June 2014)
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Identification of proton and hydrogen bond dynamics in channelrhodopsin

Principal Investigators: Prof. Dr. Joachim Heberle (FU), Prof. Dr. Karsten Heyne (FU)

In project B3, protonation dynamics that are crucial to the function of channelrhodopsin-2 (ChR2), will be monitored by time-resolved spectroscopy in the infrared and UV-visible range. The entire 15 decades of the 'chemical time-range' (10-13–102 s) will be covered to follow retinal isomerization, protein backbone alterations, changes in hydrogen-bonding and proton transfer. The project focuses on the question how light absorption by the retinal induces gating of the ion channel in ChR2 and what is the temporal sequence of the processes. Moreover, the role of specific residues and the influence of the membrane potential on the functionality of the cation channel will be addressed.

Publications 2013 - 2017

Heberle, J., Deupi, X., and Schertler, G. (2014). Retinal proteins — You can teach an old dog new tricks. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1837, 531-532.

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.]

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.

Linke, M., Yang, Y., Zienicke, B., Hammam, M.A.S., von Haimberger, T., Zacarias, A., Inomata, K., Lamparter, T., and Heyne, K. (2013). Electronic transitions and heterogeneity of the bacteriophytochrome Pr absorption band: An angle balanced polarization resolved femtosecond VIS pump-IR probe study. Biophysical Journal 105, 1756-1766.

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., and Heberle, J. (2014). Channelrhodopsin unchained: Structure and mechanism of a light-gated cation channel. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1837, 626-642.

Lórenz-Fonfría, V.A., and Heberle, J. (2014). Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy. Journal of Visualized Experiments 88, e51622. [Link to news section.]

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.

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., 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.

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.

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.

Neumann-Verhoefen, M.-K., Neumann, K., Bamann, C., Radu, I., Heberle, J., Bamberg, E., and Wachtveitl, J. (2013). Ultrafast Infrared Spectroscopy on Channelrhodopsin-2 Reveals Efficient Energy Transfer from the Retinal Chromophore to the Protein. Journal of the American Chemical Society 135, 6968-6976.

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.

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., Berg, A., Mahammed, A., Gross, Z., and Heyne, K. (2016). Ultrafast electronic and vibrational dynamics in brominated aluminum corroles: Energy relaxation and triplet formation. Structural Dynamics 3, 043210.

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.

Velazquez Escobar, F., von Stetten, D., Günther-Lütkens, M., Keidel, A., Michael, N., Lamparter, T., Essen, L.-O., Hughes, J., Gärtner, W., Yang, Y., Heyne, K., Mroginski, M.A., and Hildebrandt, P. (2015). Conformational heterogeneity of the Pfr chromophore in plant and cyanobacterial phytochromes. Frontiers in Molecular Biosciences 2, 37.

Yang, Y., Heyne, K., Mathies, R.A., and Dasgupta, J. (2016). Non-Bonded Interactions Drive the Sub-Picosecond Bilin Photoisomerization in the Pfr State of Phytochrome Cph1. ChemPhysChem 17, 369-374.

Yang, Y., Linke, M., von Haimberger, T., Matute, R., González, L., Schmieder, P., and Heyne, K. (2014). Active and silent chromophore isoforms for phytochrome Pr photoisomerization: An alternative evolutionary strategy to optimize photoreaction quantum yields. Structural Dynamics 1, 014701.