Time-resolved NMR investigation of kinase dynamics exploiting photoswitchable kinase ligands

• Applicant: Professor Dr. Harald Schwalbe
• Subject Area: Structural Biology
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 559038889

Since proteins function by changing their conformation, tracking the dynamic events occurring inside the protein is of great interest for modern structural biology. However, such studies are impeded by the fact that there are not many approaches to analyze these fast conformational rearrangements. One of the possible ways to address this challenge is time-resolved NMR spectroscopy. There are many ways to measure the dynamics by NMR spectroscopy, one of the prospective approaches is to use light irradiation to induce the desired dynamic processes on demand. In this regard, the overall aim of this proposal is to develop new photoswitchable kinase ligands as a first proof-of-concept to conduct time-resolved NMR spectroscopy using reversible switching to regulate the signalling state of protein kinase in response to the light irradiation at two wavelengths. We plan to investigate two target protein kinases – EphA2 and p38, both having high pharmaceutical significance. For both proteins we have already designed the photoswitchable ligands that can bind to kinases with high affinity and change the binding mode in response to irradiation. The compounds that were already characterized reveal an almost zero photofatigueness, allowing creating the same desired state of the system under investigation at the beginning of each scan of an NMR experiment. This makes those ligands perfect for recording two-dimensional and multidimensional NMR spectra, providing the highest possible resolution to track the protein dynamics in multiple timescales. The general intention of the project is to provide the basis to extend the time-resolved NMR studies to additional protein targets with high pharmaceutical significance.

• DFG Programme: Research Grants