Spectral and kinetic characterization of electron acceptor A1 in a Photosystem I core devoid of iron-sulfur centers F X, F B and F A.

The kinetic and spectroscopic properties of the secondary electron acceptor A1 were determined by flash absorption spectroscopy at room and cryogenic temperatures in a Photosystem I (PS I) core devoid of the iron-sulfur clusters FX, FB and FA. It was shown earlier (Warren, P.V., Golbeck, J.H. and Warden, J.T. (1993) Biochemistry 32: 849-857) that the majority of the flash-induced absorbance increase at 820 nm, reflecting formation of P700(+), decays with a t1/2 of 10 μs due to charge recombination between P700(+) and A1 (-). Following A1 (-) directly around 380 nm, where absorbance changes due to the formation of P700(+) are negligible, two major decay components were resolved in this study with t1/2 of ≈ 10 μs and 110 μs at an amplitude ratio of ≈ 2.5:1. The difference spectra between 340 and 490 nm of the two kinetic phases are highly similar, showing absorbance increases from 340 to 400 nm characteristic of the one-electron reduction of the phylloquinone A1. When measured at 10 K, the flash-induced absorbance changes around 380 nm can be fitted with two decay phases of t1/2 ≈ 15 μs and 150 μs at an amplitude ratio ≈ 1:1. The difference spectra of both kinetic phases from 340 to 400 nm are similar to those determined at 298 K and are therefore attributed to charge recombination in the pair P700(+)A1 (-). These results indicate that the backreaction between P700(+) and A1 (-) is multiphasic when FX, FB and FA are removed, and only slightly temperature dependent in the range of 298 K to 10 K.