Investigation of the Stationary and Transient A(1) Radical in Trp --> Phe Mutants of Photosystem I

• Published in: Applied magnetic resonance Vol. 38; no. 2; pp. 187 - 203
• Main Authors: Niklas, Jens, Gopta, Oxana, Epel, Boris, Lubitz, Wolfgang, Antonkine, Mikhail L
• Format: Journal Article
• Language: English
• Published: Austria 01-05-2010
• ISSN: 0937-9347
• DOI: 10.1007/s00723-009-0112-5

Photosystem I (PS I) contains two symmetric branches of electron transfer cofactors. In both the A- and B-branches, the phylloquinone in the A(1) site is pi-stacked with a tryptophan residue and is H-bonded to the backbone nitrogen of a leucine residue. In this work, we use optical and electron paramagnetic resonance (EPR) spectroscopies to investigate cyanobacterial PS I complexes, where these tryptophan residues are changed to phenylalanine. The time-resolved optical data show that backward electron transfer from the terminal electron acceptors to P(700) (.+) is affected in the A- and B-branch mutants, both at ambient and cryogenic temperatures. These results suggest that the quinones in both branches take part in electron transport at all temperatures. The electron-nuclear double resonance (ENDOR) spectra of the spin-correlated radical pair P(700) (.+)A(1) (.-) and the photoaccumulated radical anion A(1) (.-), recorded at cryogenic temperature, allowed the identification of characteristic resonances belonging to protons of the methyl group, some of the ring protons and the proton hydrogen-bonded to phylloquinone in the wild type and both mutants. Significant changes in PS I isolated from the A-branch mutant are detected, while PS I isolated from the B-branch mutant shows the spectral characteristics of wild-type PS I. A possible short-lived B-branch radical pair cannot be detected by EPR due to the available time resolution; therefore, only the A-branch quinone is observed under conditions typically employed for EPR and ENDOR spectroscopies.