Electron transfer in cyanobacterial Photosystem I. II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX

Wu Xu, Parag R. Chitnis, Alfia Valieva, Art Van der Est, Klaus Brettel, Mariana Guergova-Kuras, Yulia N. Pushkar, Stephan G. Zech, Dietmar Stehlik, Gaozhong Shen, Boris Zybailov, John H. Golbeck

Research output: Contribution to journalArticle

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Abstract

The directionality of electron transfer in Photosystem I (PS I) is investigated using site-directed mutations in the phylloquinone (QK) and Fx binding regions of Synnechocystis sp. PCC 6803. The kinetics of forward electron transfer from the secondary acceptor A1 (phylloquinone) were measured in mutants using time-resolved optical difference spectroscopy and transient EPR spectroscopy. In whole cells and PS I complexes of the wild-type both techniques reveal a major, slow kinetic component of τ ≈ 300 ns while optical data resolve an additional minor kinetic component of τ ≈ 10 ns. Whole cells and PS I complexes from the W697FPsaA and S692CPsaA mutants show a significant slowing of the slow kinetic component, whereas the W677FPsaB and S672CPsaB mutants show a less significant slowing of the fast kinetic component. Transient EPR measurements at 260 K show that the slow phase is ∼3 times slower than at room temperature. Simulations of the early time behavior of the spin polarization pattern of P700+A1-, in which the decay rate of the pattern is assumed to be negligibly small, reproduce the observed EPR spectra at 260 K during the first 100 ns following laser excitation. Thus any spin polarization from P700+Fx- in this time window is very weak. From this it is concluded that the relative amplitude of the fast phase is negligible at 260 K or its rate is much less temperature-dependent than that of the slow component. Together, the results demonstrate that the slow kinetic phase results from electron transfer from QK-A to Fx and that this accounts for at least 70% of the electrons. Although the assignment of the fast kinetic phase remains uncertain, it is not strongly temperature dependent and it represents a minor fraction of the electrons being transferred. All of the results point toward asymmetry in electron transfer, and indicate that forward transfer in cyanobacterial PS I is predominantly along the PsaA branch.

Original languageEnglish (US)
Pages (from-to)27876-27887
Number of pages12
JournalJournal of Biological Chemistry
Volume278
Issue number30
DOIs
StatePublished - Jul 25 2003

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Photosystem I Protein Complex
Photosystem II Protein Complex
Electrons
Kinetics
Vitamin K 1
Paramagnetic resonance
Spin polarization
Temperature
Spectrum Analysis
Spectroscopy
Laser excitation
Lasers
Mutation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Xu, Wu ; Chitnis, Parag R. ; Valieva, Alfia ; Van der Est, Art ; Brettel, Klaus ; Guergova-Kuras, Mariana ; Pushkar, Yulia N. ; Zech, Stephan G. ; Stehlik, Dietmar ; Shen, Gaozhong ; Zybailov, Boris ; Golbeck, John H. / Electron transfer in cyanobacterial Photosystem I. II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 30. pp. 27876-27887.
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abstract = "The directionality of electron transfer in Photosystem I (PS I) is investigated using site-directed mutations in the phylloquinone (QK) and Fx binding regions of Synnechocystis sp. PCC 6803. The kinetics of forward electron transfer from the secondary acceptor A1 (phylloquinone) were measured in mutants using time-resolved optical difference spectroscopy and transient EPR spectroscopy. In whole cells and PS I complexes of the wild-type both techniques reveal a major, slow kinetic component of τ ≈ 300 ns while optical data resolve an additional minor kinetic component of τ ≈ 10 ns. Whole cells and PS I complexes from the W697FPsaA and S692CPsaA mutants show a significant slowing of the slow kinetic component, whereas the W677FPsaB and S672CPsaB mutants show a less significant slowing of the fast kinetic component. Transient EPR measurements at 260 K show that the slow phase is ∼3 times slower than at room temperature. Simulations of the early time behavior of the spin polarization pattern of P700+A1-, in which the decay rate of the pattern is assumed to be negligibly small, reproduce the observed EPR spectra at 260 K during the first 100 ns following laser excitation. Thus any spin polarization from P700+Fx- in this time window is very weak. From this it is concluded that the relative amplitude of the fast phase is negligible at 260 K or its rate is much less temperature-dependent than that of the slow component. Together, the results demonstrate that the slow kinetic phase results from electron transfer from QK-A to Fx and that this accounts for at least 70{\%} of the electrons. Although the assignment of the fast kinetic phase remains uncertain, it is not strongly temperature dependent and it represents a minor fraction of the electrons being transferred. All of the results point toward asymmetry in electron transfer, and indicate that forward transfer in cyanobacterial PS I is predominantly along the PsaA branch.",
author = "Wu Xu and Chitnis, {Parag R.} and Alfia Valieva and {Van der Est}, Art and Klaus Brettel and Mariana Guergova-Kuras and Pushkar, {Yulia N.} and Zech, {Stephan G.} and Dietmar Stehlik and Gaozhong Shen and Boris Zybailov and Golbeck, {John H.}",
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Electron transfer in cyanobacterial Photosystem I. II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX. / Xu, Wu; Chitnis, Parag R.; Valieva, Alfia; Van der Est, Art; Brettel, Klaus; Guergova-Kuras, Mariana; Pushkar, Yulia N.; Zech, Stephan G.; Stehlik, Dietmar; Shen, Gaozhong; Zybailov, Boris; Golbeck, John H.

In: Journal of Biological Chemistry, Vol. 278, No. 30, 25.07.2003, p. 27876-27887.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Electron transfer in cyanobacterial Photosystem I. II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX

AU - Xu, Wu

AU - Chitnis, Parag R.

AU - Valieva, Alfia

AU - Van der Est, Art

AU - Brettel, Klaus

AU - Guergova-Kuras, Mariana

AU - Pushkar, Yulia N.

AU - Zech, Stephan G.

AU - Stehlik, Dietmar

AU - Shen, Gaozhong

AU - Zybailov, Boris

AU - Golbeck, John H.

PY - 2003/7/25

Y1 - 2003/7/25

N2 - The directionality of electron transfer in Photosystem I (PS I) is investigated using site-directed mutations in the phylloquinone (QK) and Fx binding regions of Synnechocystis sp. PCC 6803. The kinetics of forward electron transfer from the secondary acceptor A1 (phylloquinone) were measured in mutants using time-resolved optical difference spectroscopy and transient EPR spectroscopy. In whole cells and PS I complexes of the wild-type both techniques reveal a major, slow kinetic component of τ ≈ 300 ns while optical data resolve an additional minor kinetic component of τ ≈ 10 ns. Whole cells and PS I complexes from the W697FPsaA and S692CPsaA mutants show a significant slowing of the slow kinetic component, whereas the W677FPsaB and S672CPsaB mutants show a less significant slowing of the fast kinetic component. Transient EPR measurements at 260 K show that the slow phase is ∼3 times slower than at room temperature. Simulations of the early time behavior of the spin polarization pattern of P700+A1-, in which the decay rate of the pattern is assumed to be negligibly small, reproduce the observed EPR spectra at 260 K during the first 100 ns following laser excitation. Thus any spin polarization from P700+Fx- in this time window is very weak. From this it is concluded that the relative amplitude of the fast phase is negligible at 260 K or its rate is much less temperature-dependent than that of the slow component. Together, the results demonstrate that the slow kinetic phase results from electron transfer from QK-A to Fx and that this accounts for at least 70% of the electrons. Although the assignment of the fast kinetic phase remains uncertain, it is not strongly temperature dependent and it represents a minor fraction of the electrons being transferred. All of the results point toward asymmetry in electron transfer, and indicate that forward transfer in cyanobacterial PS I is predominantly along the PsaA branch.

AB - The directionality of electron transfer in Photosystem I (PS I) is investigated using site-directed mutations in the phylloquinone (QK) and Fx binding regions of Synnechocystis sp. PCC 6803. The kinetics of forward electron transfer from the secondary acceptor A1 (phylloquinone) were measured in mutants using time-resolved optical difference spectroscopy and transient EPR spectroscopy. In whole cells and PS I complexes of the wild-type both techniques reveal a major, slow kinetic component of τ ≈ 300 ns while optical data resolve an additional minor kinetic component of τ ≈ 10 ns. Whole cells and PS I complexes from the W697FPsaA and S692CPsaA mutants show a significant slowing of the slow kinetic component, whereas the W677FPsaB and S672CPsaB mutants show a less significant slowing of the fast kinetic component. Transient EPR measurements at 260 K show that the slow phase is ∼3 times slower than at room temperature. Simulations of the early time behavior of the spin polarization pattern of P700+A1-, in which the decay rate of the pattern is assumed to be negligibly small, reproduce the observed EPR spectra at 260 K during the first 100 ns following laser excitation. Thus any spin polarization from P700+Fx- in this time window is very weak. From this it is concluded that the relative amplitude of the fast phase is negligible at 260 K or its rate is much less temperature-dependent than that of the slow component. Together, the results demonstrate that the slow kinetic phase results from electron transfer from QK-A to Fx and that this accounts for at least 70% of the electrons. Although the assignment of the fast kinetic phase remains uncertain, it is not strongly temperature dependent and it represents a minor fraction of the electrons being transferred. All of the results point toward asymmetry in electron transfer, and indicate that forward transfer in cyanobacterial PS I is predominantly along the PsaA branch.

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