EPR of Type I photosynthetic reaction centers

Light-induced reactions in photosynthetic reaction centers are initiated by the absorption of a photon, which results in the transfer of a single electron and the generation of radical ions in the donor and acceptor molecules involved in the charge-separated state. Electron paramagnetic resonance (EPR) spectroscopy is the ideal method for the study of such reactions. In addition to measuring spectra of the electron transfer cofactors in continuous light, reactions can be initiated by brief flashes of light, thereby allowing the kinetics of forward electron transfer as well as recombination reactions to be obtained. Because the donor and acceptor pairs are so closely spaced and because light induced charge separation is so rapid, the donor and early acceptors are in a quantum mechanically spin entangled state, which confers properties such as increased sensitivity, the ability to measure reactions on the nanosecond timescale, and the determination of bond angles between cofactors. Additionally, distances between pairs of cofactors can be measured by detecting the modulation of a phase shifted “out-of-phase” electron spin echo signal. In this methods article, we will describe how continuous wave EPR, time resolved EPR, and pulsed EPR can be used to measure these properties in Type I photosynthetic reaction centers. Methods of analysis are described for the bound electron transfer cofactors in the heterodimeric Photosystem I reaction center of plants and cyanobacteria and in the homodimeric reaction centers found in phototrophic members of the phyla Bacillota, Chlorobiota, and Acidobacteriota.