title: Incorporation of a high potential quinone reveals that electron transfer in Photosystem I becomes highly asymmetric at low temperature.
abstract: Photosystem I (PS I) has two nearly identical branches of electron-transfer co-factors. Based on point mutation studies, there is general agreement that both branches are active at ambient temperature but that the majority of electron-transfer events occur in the A-branch. At low temperature, reversible electron transfer between P(700) and A(1A) occurs in the A-branch. However, it has been postulated that irreversible electron transfer from P(700) through A(1B) to the terminal iron-sulfur clusters F(A) and F(B) occurs via the B-branch. Thus, to study the directionality of electron transfer at low temperature, electron transfer to the iron-sulfur clusters must be blocked. Because the geometries of the donor-acceptor radical pairs formed by electron transfer in the A- and B-branch differ, they have different spin-polarized EPR spectra and echo-modulation decay curves. Hence, time-resolved, multiple-frequency EPR spectroscopy, both in the direct-detection and pulse mode, can be used to probe the use of the two branches if electron transfer to the iron-sulfur clusters is blocked. Here, we use the PS I variant from the menB deletion mutant strain of Synechocyctis sp. PCC 6803, which is unable to synthesize phylloquinone, to incorporate 2,3-dichloro-1,4-naphthoquinone (Cl(2)NQ) into the A(1A) and A(1B) binding sites. The reduction midpoint potential of Cl(2)NQ is approximately 400 mV more positive than that of phylloquinone and is unable to transfer electrons to the iron-sulfur clusters. In contrast to previous studies, in which the iron-sulfur clusters were chemically reduced and/or point mutations were used to prevent electron transfer past the quinones, we find no evidence for radical-pair formation in the B-branch. The implications of this result for the directionality of electron transfer in PS I are discussed.
authors.affiliation: Department of Chemistry, Brock University, St. Catharines, ON, Canada.
authors.forename: Sam
authors.initials: S
authors.lastname: Mula
authors.forename: Anton
authors.initials: A
authors.lastname: Savitsky
authors.forename: Klaus
authors.initials: K
authors.lastname: Möbius
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.forename: John H
authors.initials: JH
authors.lastname: Golbeck
authors.forename: Mahir D
authors.initials: MD
authors.lastname: Mamedov
authors.forename: Alexey Yu
authors.initials: AY
authors.lastname: Semenov
authors.forename: Art
authors.initials: A
authors.lastname: van der Est
mesh_terms: D001665:Binding Sites; D003080:Cold Temperature; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D007700:Kinetics; D009154:Mutation; D009285:Naphthoquinones / Q000737:chemistry; D045331:Photosystem I Protein Complex / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011809:Quinones / Q000737:chemistry*; D013696:Temperature
chemical_list: D007506:Iron-Sulfur Proteins; D009285:Naphthoquinones; D045331:Photosystem I Protein Complex; D011809:Quinones; C007332:dichlone