Technology research and method development for studying protein single crystals of nanoliter volumes (0.2 mm3) at X-band using advanced resonators and state-of-the-art digital detection.
Electron Paramagnetic Resonance (EPR) experiments on single crystals are the ultimate method to study paramagnetic states and obtain the full magnetic tensor interactions reflecting the electronic structure of the active site. Ultimately the catalytic activity of hydrogenase enzymes can be understood by relating the information of the magnetic principal axes to the known protein structure. However, the application of single-crystal EPR is severely limited by the small crystals sizes that are usually available (sub-nanoliter to nanoliter volumes). We propose the following work packages to make single crystal EPR feasible.
In this work package, we introduce a self-resonant micro-helix with an inner diameter of 0.4 mm. With this geometry the microwave efficiency parameter up to 3.5 mT/W1/2 are possible.
This work package explores the usage of Non-adiabatic Rapid Scan (NARS) and Arbitrary Waveform Generators (AWGs) to enhance signal-to-noise by modern digital detection.
This work package makes use of the key enabling technologies in order to study, for the first time, protein single crystals of [FeFe]-Hydrogenase enzymes.
The Key Enabling Technologies outlined in this fellowship have the potential to increase the sensitivity of EPR by a factor of 30 through the application of highly innovative concepts based on micro-resonators.
