Role of pheophytin alpha in the primary charge separation of photosystem I from Acaryochloris marina: Femtosecond optical studies of excitation energy and electron transfer reactions

Photosystem I (PSI) of the cyanobacterium Acaryochloris marina is capable of performing an efficient photo-electrochemical conversion of far-red light due to its unique suite of cofactors. Chlorophyll d (Chl-d) has been long known as the major antenna pigment in the PSI from A. marina, while the exact cofactor composition of the reaction centre (RC) was established only recently by cryo-electron microscopy. The RC consists of four Chld molecules, and, surprisingly, two molecules of pheophytin alpha (Pheo-alpha), which provide a unique opportunity to resolve, spectrally and kinetically, the primary electron transfer reactions. Femtosecond transient absorption spectroscopy was here employed to observe absorption changes in the 400-860 nm spectral window occurring in the 0.1-500 ps timescale upon unselective antenna excitation and selective excitation of the Chl-d special pair P-740 in the RC. A numerical decomposition of the absorption changes, including principal component analysis, allowed the identification of P-740((+)) Chl(d2)((-)) as the primary charge separated state and P-740((+)) Pheo(alpha 3)((-)) as the successive, secondary, radical pair. A remarkable feature of the electron transfer reaction between Chld2 and Pheoa3 is the fast, kinetically unresolved, equilibrium with an estimated ratio of 1:3. The energy level of the stabilised ionradical state P-740((+)) Pheo(alpha 3)((-)) was determined to be similar to 60 meV below that of the RC excited state. In this regard, the energetics and the structural implications of the presence of Pheo-a in the electron transfer chain of PSI from A. marina are discussed, also in comparison with those of the most diffused Chl-alpha binding RC.