Photosystem I charge separation in the absence of centers A and B. I. Optical characterization of center 'A₂' and evidence for its association with a 64-kDa peptide

The flash-induced absorption transient at 698 nm in a Photosystem I subchloroplast particle showed the following characteristics after addition of 0.25-2.0% lithium dodecyl sulfate (LDS). (i) The 30-ms transient corresponding to the P-700⁺ P-430^- backreaction was replaced by a 1.2-ms transient. (ii) The amplitude of the transient did not change immediately after LDS addition, but decayed with a half-life of 10 min at pH 8.5. (iii) Methyl viologen had no effect on the magnitude or kinetics of the transient, indicating that it cannot accept an electron from this component. (iv) The difference spectrum of the transient from 400 nm to 500 nm was characteristic of an iron-sulfur protein. (v) The transient followed first-order Arrhenius behavior between 298 K and 225 K with an activation energy of 13.3 kJ/mol; between 225 K and 77 K, the 85-ms half-time remained temperature-invariant. These properties suggest that the LDS-induced absorption transient corresponds to the P-700⁺ A^-₂ change recombination seen in the absence of a reduced electron-acceptor system. In the presence of LDS, the reaction-center complex was dissociated, allowing removal of the smaller peptides from the 64-kDa P-700-containing protein. With prolonged incubation, the iron-sulfur clusters were destroyed through conversion of the labile sulfide to zero-valence sulfur. About 35% of the zero-valence sulfur was found associated with the 64-kDa protein under conditions that allowed separation of the small peptides. We interpret the long lifetime of the P-700⁺ A^-₂ transient after LDS addition and the association of zero-valence sulfur with a 64-kDa protein to indicate that A₂ is closely associated with, and perhaps integral with, the P-700-containing protein.