Pulsed EPR spectroscopy on short-lived intermediates in Photosystem I.

The application of pulsed electron paramagnetic resonance spectroscopy on short-lived intermediates in Photosystem I is reviewed. The spin polarization in light-induced radical pairs gives rise to a phase shifted 'out-of-phase' electron spin echo signal. This echo signal shows a prominent modulation of its intensity as a function of the spacing between the two microwave pulses. Its modulation frequency is determined by the electron-electron spin couplings within the radical pair. Thereby, the measurement of the dipolar coupling gives direct information about the spin-spin distance and can therefore be used to determine cofactor distances with high precision. Application of this technique to the radical pair P(*+)(700)A(*-)(1) in Photosystem I is discussed. Moreover, if oriented samples (e.g. single crystals) are used, the angular dependence of the dipolar coupling can be used to derive the orientation of the axis connecting donor and acceptor with respect to an external (crystal) axes system. Using out-of-phase electron spin echo envelope modulation spectroscopy, the localization of the secondary acceptor quinone A(1) has become possible.