Dissecting the mechanism of proton and cation conductance in viroporins using vibrational spectroscopy

Principal Investigators:  Dr. Jacek Kozuch (FUB)

In this project, we will use infrared spectroscopic methods to provide insight into the mechanisms of proton and cation conductance of the viroporins of the mosquito-borne Dengue 1 and the West Nile viruses. We will identify key amino acid residues that may be responsible for pH sensing or gating as well as monitor changes in structure and electrostatics of the channel interior, the latter using vibrational Stark effect probes, associated with the pore opening process. Finally, we will test the influence of viroporin inhibitors on the conductance mechanism and evaluate potential antiviral-resistance conferring mutations.

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. doi: 10.3389/fchem.2021.669452.

Kozuch, J., Schneider, S. H., Zheng, C., Ji, Z., Bradshaw, R.T., and Boxer, S.G. (2021). Testing the limitations of MD-based local electric fields using the vibrational Stark effect in solution: pencillin G as a test case. J Phys Chem B, 125: 4415-4427. doi: 10.1021/acs.jpcb.1c00578.

Kraskov, A., von Sass, J., Nguyen, A.D., Hoang, T.O., Buhrke, D., Katz, S., Michael, N., Kozuch, J., Zebger, I., Siebert, F., Scheerer, P., Mroginski, M.A., Budisa, N., and Hildebrandt P. (2021). Local Electric Field Changes during the Photoconversion of the Bathy Phytochrome Agp2. Biochemistry, 60, 40, 2967-2977. doi: 10.1021/acs.biochem.1c00426.

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.

Paschke, R. R., Mohr, S., Lange, S., Lange, A., Kozuch, J. (2023). In situ spectroscopic detection of large-scale reorientations of transmembrane helices during influenza A M2 channel opening. Angew Chem Int Ed, 62, e202309069.
doi: 10.1002/anie.202309069.

Weaver, J.B., Kozuch, J., Kirsh, J.M. and Boxer, S.G. (2022). Nitrile Infrared Intensities Characterize Electric Fields and Hydrogen Bonding in Protic, Aprotic, and Protein Environments. J Am Chem Soc, 144, 17: 7562-7567. doi: 10.1021/jacs.2c00675

Zheng, C., Mao, Y., Kozuch, J., Atsango, A.O., Ji, Z., Markland, T.E. and Boxer, S.G. (2022). A two-directional vibrational probe reveals different electric field orientations in solution and an enzyme active site. Nat Chem. doi: 10.1038/s41557-022-00937-w.

Kirsh, J.M., Weaver, J.B., Boxer, S.G. and Kozuch, J. (2024). Critical Evaluation of Polarizable and Nonpolarizable Force Fields for Proteins Using Experimentally Derived Nitrile Electric Fields. J Am Chem Soc, 146, 10: 6983-6991. doi: 10.1021/jacs.3c14775.