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Project B1

Proton exchange processes in phytochromes and retinal proteins investigated by MAS NMR

Principal Investigators: Prof. Dr. Hartmut Oschkinat (FMP)

Protonation dynamics associated with structural changes in phytochromes are investigated by very fast magic-angle-spinning (MAS) NMR, detecting exchangeable protons directly. Proton exchange pattern within the chromophore and with residues towards the tongue region between the Pr, Meta-F, and Pfr states of the bacteriophytochromes Agp2 (batho), Agp1 (prototypic), and Cph1 will be analyzed. In this context, the proposal of a keto-enol tautomerism in the biliverdin of Agp2 will be scrutinized including a derivation of kinetic parameters. Furthermore, we will localize exchangeable protons in the channels of channelrhodopsins C1C2 and iC++slow and study their exchange pattern.


2017 - 2020

Friedrich, D., Brünig, F.N., Nieuwkoop, A.J., Netz, R., Hegemann, P. and Oschkinat, H. (2020). Collective exchange processes reveal an active site proton cage in bacteriorhodopsin. Commun. Biol. 3, doi: 10.1038/s42003-019-0733-7.

Friedrich, D., Perodeau, J., Nieuwkoop, A.J., and Oschkinat, H. (2020). MAS NMR detection of hydrogen bonds for protein secondary structure characterization. J. Biomol. NMR 74(4-5):247-256; doi: 10.1007/s10858-020-00307-z

Fudim, R., Szczepek, M., Vierock, J., Vogt, A., Schmidt, A., Kleinau, G., Fischer, P., Bartl, F., Scheerer, P., Hegemann, P. (2019). Design of a light-gated proton channel based on the crystal structure of Coccomyxa rhodopsin. Sci Signal. 2019 Mar 19;12(573). pii: eaav4203. doi: 10.1126/scisignal.aav4203.

Geiger, M.-A., Jagtap, A.P., Kaushik, M., Sun, H., Stöppler, D., Sigurdsson, S.T., Corzilius, B., and Oschkinat, H. (2018). Efficiency of Water‐Soluble Nitroxide Biradicals for Dynamic Nuclear Polarization in Rotating Solids at 9.4 T: bcTol‐M and cyolyl‐TOTAPOL as New Polarizing Agents. Chemistry 24(51), doi: 10.1002/chem.201801251.

Gerland, L.M., Friedrich, D., Hopf, L., Donovan, E., Wallmann, A., Erdmann, N., Diehl, A., Bommer, M., Buzar, K., Ibrahim, M., Schmieder, P., Dobbek, H., Zouni, A., Bondar, A.-N., Dau, H., and Oschkinat, H. (2020). pH‐Dependent Protonation of Surface Carboxylates in PsbO Enables Local Buffering and Triggers Structural Changes. ChemBioChem 21, 1597-1604. doi: 10.1002/cbic.201900739.

Grimm, C., Silapetere, A., Vogt, A., Bernal S., Yinth A., and Hegemann, P. (2018). Electrical properties, substrate specificity and optogenetic potential of the engineered light-driven sodium pump eKR2. Sci Rep 8, 9316; doi: 10.1038/s41598-018-27690-w

Krause, B.S., Grimm, C., Kaufmann, J.C.D., Schneider, F., Sakmar, T.P., and Hegemann, P. (2017). Complex photochemistry within the green-absorbing channelrhodopsin ReaChR. Biophys. J. 112, 1166-1175; doi: 10.1016/j.bpj.2017.02.001

Krause, B.S., Kaufmann, J.C.D., Kuhne, J., Vierock, V., Huber, T., Sakmar, T.P., Gerwert, K., Bartl, F.J., and Hegemann, P. (2019). Tracking pore hydration in channelrhodopsin by site-directed infraredactive azido probes. Biochemistry 58, 1275-1286; doi: 10.1021/acs.biochem.8b01211

Mukherjee, S., Hegemann, P., Broser, M. (2019). Enzymerhodopsins: novel photoregulated catalysts for optogenetics. Curr Opin Struct Biol. 57:118-126. doi: 10.1016/j.sbi.2019.02.003.

Oda, K., Vierock, J., Oishi, S., Rodriguez-Rozada, S., Taniguchi, R., Yamashita, K., Wiegert, J.S., Nishizawa, T., Hegemann, P., and Nureki, O. (2018). Crystal structure of the red light-activated channelrhodopsin Chrimson. Nat. Commun. 9:3949; doi: 10.1038/s41467-018-06421-9

Scheib, U., Broser, M., Constantin, O. M., Yang, S.,Gao, S., Mukherjee, S., Stehfest, K., Nagel, G., Gee, C. E., and Hegemann, P. (2018). Rhodopsin-cyclases for photocontrol of cGMP/cAMP and 2.3 Å structure of the adenylyl cyclase domain. Nature Commun 9, 2046.; doi: 10.1038/s41467-018-04428-w

Stöppler. D., Macpherson. A., Smith-Penzel, S., Basse, N., Lecomte, F., Deboves, H., Taylor, R.D., Norman, T., Porter, J., Waters, L.C., Westwood, M., Cossins, B., Cain, K., White, J., Griffin, R., Prosser, C., Kelm, S., Sullivan, A.H., Fox, D. 3rd, Carr, M.D., Henry, A., Taylor, R., Meier, B.H., Oschkinat, H., and Lawson, A.D. (2018). Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR. PLoS Biol. 16(5), doi: 10.1371/journal.pbio.2006192

Vierock, J., Grimm, C., Nitzen, N., and Hegemann, P. (2017). Molecular determinants of proton selectivity and gating in the red-light activated channelrhodopsin Chrimson. Sci. Rep. 7, 9928; doi: 10.1038/s41598-017-09600-8

2013 - 2016

Bruun, S., Stoeppler, D., Keidel, A., Kuhlmann, U., Luck, M., Diehl, A., Geiger, M.-A., Woodmansee, D., Trauner, D., Hegemann, P., Oschkinat, H., Hildebrandt, P., and Stehfest, K. (2015). Light–Dark Adaptation of Channelrhodopsin Involves Photoconversion between the all-trans and 13-cis Retinal Isomers. Biochemistry 54, 5389-5400.

Ernst, O.P., Lodowski, D.T., Elstner, M., Hegemann, P., Brown, L.S., and Kandori, H. (2014). Microbial and Animal Rhodopsins: Structures, Functions, and Molecular Mechanisms. Chemical Reviews 114, 126-163.

Jagtap, A.P., Geiger, M.-A., Stöppler, D., Orwick-Rydmark, M., Oschkinat, H., and Sigurdsson, S.T. (2016). bcTol: A highly water-soluble biradical for efficient dynamic nuclear polarization of biomolecules. Chemical Communications 52, 7020-7023.

Kuhne, J., Eisenhauer, K., Ritter, E., Hegemann, P., Gerwert, K., and Bartl, F. (2015). Early Formation of the Ion-Conducting Pore in Channelrhodopsin-2. Angewandte Chemie International Edition 54, 4953-4957.

Luck, M., Bruun, S., Keidel, A., Hegemann, P., and Hildebrandt, P. (2015). Photochemical chromophore isomerization in histidine kinase rhodopsin HKR1. FEBS Letters 589, 1067-1071.

Ritter, E., Piwowarski, P., Hegemann, P., and Bartl, F.J. (2013). Light-dark Adaptation of Channelrhodopsin C128T Mutant. Journal of Biological Chemistry 288, 10451-10458.

Scheib, U., Stehfest, K., Gee, C.E., Korschen, H.G., Fudim, R., Oertner, T.G., and Hegemann, P. (2015). The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella emersonii enables fast optical control of cGMP signaling. Science Signaling 8, rs8.

Stöppler, D., Song, C., van Rossum, B.-J., Geiger, M.-A., Lang, C., Mroginski, M.-A., Jagtap, A.P., Sigurdsson, S.T., Matysik, J., Hughes, J., and Oschkinat, H. (2016). Dynamic nuclear polarization provides new insights into chromophore structure in phytochrome photoreceptors. Angew Chem Int Ed 55, 16017–16020, doi: doi: 10.1002/anie.201608119.