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Consequences of structural modifications in cytochrome b559 on the electron acceptor side of Photosystem II

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Abstract

Cytb559 in Photosystem II is a heterodimeric b-type cytochrome. The subunits, PsbE and PsbF, consist each in a membrane α-helix. Mutants were previously designed and studied in Thermosynechococcus elongatus (Sugiura et al., Biochim Biophys Acta 1847:276–285, 2015) either in which an axial histidine ligand of the haem-iron was substituted for a methionine, the PsbE/H23M mutant in which the haem was lacking, or in which the haem environment was modified, the PsbE/Y19F and PsbE/T26P mutants. All these mutants remained active showing that the haem has no structural role provided that PsbE and PsbF subunits are present. Here, we have carried on the characterization of these mutants. The following results were obtained: (i) the Y19F mutation hardly affect the Em of Cytb559, whereas the T26P mutation converts the haem into a form with a Em much below 0 mV (so low that it is likely not reducible by QB−) even in an active enzyme; (ii) in the PsbE/H23M mutant, and to a less extent in PsbE/T26P mutant, the electron transfer efficiency from QA− to QB is decreased; (iii) the lower Em of the QA/QA− couple in the PsbE/H23M mutant correlates with a higher production of singlet oxygen; (iv) the superoxide and/or hydroperoxide formation was not increased in the PsbE/H23M mutant lacking the haem, whereas it was significantly larger in the PsbE/T26P. These data are discussed in view of the literature to discriminate between structural and redox roles for the haem of Cytb559 in the production of reactive oxygen species.

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Abbreviations

Chl:

Chlorophyll

ChlD1/ChlD2 :

Accessory Chl on the D1 or D2 side, respectively

Cyt:

Cytochrome

DCBQ:

2,6-Dichloro-p-benzoquinone

DCMU:

3-(3,4-Dichlorophenyl)-1,1-dimethylurea

E m :

Midpoint redox potential versus SHE, the standard hydrogen electrode

HP, IP, LP forms:

High-potential, intermediate-potential and low-potential forms of Cytb559

MES:

2–(N–morpholino) ethanesulfonic acid

P680 :

Primary electron donor

PD1 and PD2 :

Chl monomer of P680 on the D1 or D2 side, respectively

Pheo:

Pheophytin

PSII:

Photosystem II

P680 :

Chlorophyll dimer acting as the electron donor

4-POBN:

α-(4-Pyridyl N-oxide)-N-tert-butylnitrone

QA :

Primary quinone acceptor

QB :

Secondary quinone acceptor

T. elongatus :

Thermosynechococcus elongatus

ROS:

Reactive oxygen species

TL:

Thermoluminescence

WT*3:

T. elongatus mutant strain containing only the psbA3 gene

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Acknowledgements

We would like to thank Anja Krieger-Liszkay (CEA Saclay) for discussions and technical suggestions for the trapping of ROS. This work was supported by JSPS-KAKENHI grant in Scientific Research on Innovative Areas “Innovations for Light-Energy Conversion (I4LEC)” (17H064351 for M. S.) and a JSPS-KAKENHI grant (17K07367 for M.S.). AB was supported in part by the French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INBS-05.

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Authors and Affiliations

  1. Graduate School of Science and Technology, Ehime University, Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan

    Makoto Nakamura & Miwa Sugiura

  2. I2BC, CNRS UMR 9198, CEA Saclay, 91191, Gif-sur-Yvette, France

    Alain Boussac

  3. Proteo-Science Research Center, Ehime University, Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan

    Miwa Sugiura

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  1. Makoto Nakamura
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Correspondence to Miwa Sugiura.

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Nakamura, M., Boussac, A. & Sugiura, M. Consequences of structural modifications in cytochrome b559 on the electron acceptor side of Photosystem II. Photosynth Res 139, 475–486 (2019). https://doi.org/10.1007/s11120-018-0521-0

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