Removal of the PsaF polypeptide biases electron transfer in favor of the PsaB branch of cofactors in Triton X-100 photosystem I complexes from Synechococcus sp. PCC 7002.
Continuous wave (CW) and transient electron paramagnetic resonance studies have implied that when PsaF is removed genetically, the double reduction of A1A is facile, the lifetime of A1A(-) is shorter and the ratio of fast to slow kinetic phases increases in PS I complexes isolated with Triton X-100 (Van der Est, A., A. I. Valieva, Y. E. Kandrashkin, G. Shen, D. A. Bryant and J. H. Golbeck [2004] Biochemistry43, 1264-1275). Changes in the lifetimes of A1A(-) and A1B(-) are characteristic of mutants involving the quinone binding sites, but changes in the relative amplitudes of A1A(-) and A1B(-) are characteristic of mutants involving the primary electron acceptors, A0A and A0B. Here, we measured the fast and slow phases of electron transfer from A1B(-) and A1A(-) to FX in psaF and psaE psaF null mutants using time-resolved CW and pump-probe optical absorption spectroscopy. The lifetime of the fast kinetic phase was found to be unaltered, but the lifetime of the slow kinetic phase was shorter in the psaF null mutant and even more so in the psaE psaF null mutant. Concomitantly, the amplitude of the fast kinetic phase increased by a factor of 1.8 and 2.0 in the psaF and psaE psaF null mutants, respectively, at the expense of the slow kinetic phase. The change in ratio of the fast to slow kinetic phases is explained as either a redirection of electron transfer through A1B at the expense of A1A, or a shortening of the lifetime of A1A(-) to become identical to that of A1B(-). The constant lifetime and the characteristics of the near-UV spectrum of the fast kinetic phase favor the former explanation. A unified hypothesis is presented of a displacement of the A-jk1 alpha-helix and switchback loop, which would weaken the H-bond from Leu722 to A1A, accounting for the acceleration of the slow kinetic phase, as well as weaken the H-bond from Tyr696 to A0A, accounting for the bias of electron transfer in favor of the PsaB branch of cofactors.
Agalarov
Rufat
R
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
Byrdin
Martin
M
Rappaport
Fabrice
F
Shen
Gaozhong
G
Bryant
Donald A
DA
van der Est
Art
A
Golbeck
John H
JH
D018360:Crystallography, X-Ray; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D017830:Octoxynol / Q000737:chemistry*; D010084:Oxidation-Reduction; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D046940:Synechococcus / Q000201:enzymology* / Q000235:genetics; D013997:Time Factors
D045331:Photosystem I Protein Complex; D017830:Octoxynol