In cell EPR of native paramagnetic protein cofactors

• Applicant: Professor Dr. Robert Bittl
• Subject Area: Analytical Chemistry
• Term: from 2012 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 221264732

Goal of this project within the priority program New Frontiers in Sensitivity for EPR Spectroscopy: From Biological Cells to Nano Materials (SPP1601) is widening the applicability of electron paramagnetic resonance (EPR) spectroscopy under in cell conditions. Among the group of projects devoted to in cell EPR, we focus on proteins with naturally occurring paramagnetic organic cofactors and metal centers and have chosen two classes of proteins as examples. These are blue-light-activated proteins (cryptochrome, LOV, BLUF) and oxygen-tolerant hydrogenases. The long term goal of the studies on the blue-light-activated proteins is understanding the influence of small molecular weight cellular components occurring at high concentrations in cells on protein function. With the work on the hydrogenases we aim at an understanding of the origin of their exceptional oxygen tolerance, in particular via the analysis of subcomplexes and maturation intermediates, which are rather unstable as isolated proteins, but rather stable under cellular conditions. These biophysical studies will be flanked by instrumentation oriented work on resonance structures specifically suited to in cell EPR. This will be largely performed in tight collaboration with the project of D. Suter in the SPP1601 devoted to microresonator development. The long term goal thereby is to push in cell EPR to the limit of single-cell detection.

• DFG Programme: Priority Programmes
• Subproject of: SPP 1601: New Frontiers in Sensitivity for EPR Spectroscopy: from Biological Cells to Nano Materials
• Co-Investigator: Dr. Christian Teutloff