CHARGE RECOMBINATION BETWEEN P700+ AND A1- OCCURS DIRECTLY TO THE GROUND-STATE OF P700 IN A PHOTOSYSTEM-I CORE DEVOID OF FX, FB, AND FA

The charge recombination between P700+ and electron acceptor A1- was studied by flash kinetic spectroscopy in a photosystem I core devoid of iron-sulfur centers F(X), F(B), and F(A). We showed previously that the majority of the flash-induced absorption change at 820 nm decayed with a 10-mus half-time, which we assigned to the disappearance of the P700 triplet formed from the backreaction of P700+ with A1-[Warren, P. V., Parrett, K. G., Warden, J. T., & Golbeck, J. H. (1990) Biochemistry 29, 6545-6550]. We have reinvestigated this assignment in the near-UV, blue, and near-IR wavelength regions. The difference spectrum from 380 to 480 nm and from 720 to 910 nm shows that the P700+ A1- charge recombination is dominated by the P700 cation rather than the P700 triplet. Accordingly, the 10-mus kinetic transient represents the direct backreaction of P700+ with A1-, which repopulates the ground state of P700. This is unlike a P700-F(A)/F(B) complex where, in the presence of reduced F(X)-, F(B)-, and F(A)-, the P700+ A1- charge recombination populates the P700 triplet state [Setif, P., & Bottin, H. (1989) Biochemistry 28, 2689-2697]. The A1 acceptor is highly susceptible to disruption by detergents in the absence of iron-sulfur center F(X). The addition of 0.1% Triton X-100 to the P700-A1 core leads to a approximately 2.5-fold increase in the magnitude of the flash-induced absorption change at 780 nm; thereafter, 85% of the absorption change decays with a 25-ns half-time and 15% decays with a 3-mus half-time. The spectrum of the 25-ns phase is a convolution of contributions from both P700+ and A0-. When the P700+ spectrum is subtracted, the spectrum of A0-displays a maximum at 760 nm and doublet minima at 412 and 438 nm and has an extinction coefficient 1.4 and 1.8 times that of P700+ at 438 and 790 nm, respectively. The spectrum of the 3-mus kinetic phase shows a broad absorption increase between 730 and 820 nm accompanied by a broad bleaching between 390 and 450 nm, consistent with the decay of the P700 triplet formed in low quantum yield from the backreaction of P700+ with A0-. The rise time of the P700 triplet was measured to be approximately 25 ns, a value identical to that of the P700+ A0- charge recombination.