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Photosynthesis Research

, Volume 139, Issue 1–3, pp 267–279 | Cite as

Site-directed mutagenesis of two amino acid residues in cytochrome b559 α subunit that interact with a phosphatidylglycerol molecule (PG772) induces quinone-dependent inhibition of photosystem II activity

  • Kaichiro Endo
  • Koichi Kobayashi
  • Hsing-Ting Wang
  • Hsiu-An Chu
  • Jian-Ren Shen
  • Hajime WadaEmail author
Original article

Abstract

X-ray crystallographic analysis (1.9-Å resolution) of the cyanobacterial photosystem II (PSII) dimer showed the presence of five phosphatidylglycerol (PG) molecules per reaction center. One of the PG molecules, PG772, is located in the vicinity of the QB-binding site. To investigate the role of PG772 in PSII, we performed site-directed mutagenesis in the cytochrome (Cyt) b559 α subunit of Synechocystis sp. PCC 6803 to change two amino acids, Thr-5 and Ser-11, which interact with PG772. The photosynthetic activity of intact cells was slightly lower in all mutants than that of cells in the control strain; however, the oxygen-evolving PSII activity was decreased markedly in cells of mutants, as measured using artificial quinones (such as p-benzoquinone). Furthermore, electron transport from QA to QB was inhibited in mutants incubated with quinones, particularly under high-intensity light conditions. Lipid analysis of purified PSII showed approximately one PG molecule per reaction center, presumably PG772, was lost in the PSII dimer from the T5A and S11A mutants compared with that in the PSII dimer from the control strain. In addition, protein analysis of monomer and dimer showed decreased levels of PsbV and PsbU extrinsic proteins in the PSII monomer purified from T5A and S11A mutants. These results suggest that site-directed mutagenesis of Thr-5 and Ser-11, which presumably causes the loss of PG772, induces quinone-dependent inhibition of PSII activity under high-intensity light conditions and destabilizes the binding of extrinsic proteins to PSII.

Keywords

Cytochrome b559 Phosphatidylglycerol Photosystem II Site-directed mutagenesis 

Abbreviations

BQ

p-Benzoquinone

BN

Blue native

Chl

Chlorophyll

Cyt

Cytochrome

DCBQ

2,6-Dichloro-p-benzoquinone

DCMU

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

DGDG

Digalactosyldiacylglycerol

DM

n-Dodecyl-β-d-maltoside

DMBQ

2,6-Dimethyl-p-benzoquinone

Em

Erythromycin

EmR

Erythromycin-resistant gene cassette

Fecy

Potassium ferricyanide

HP

High potential

IP

Intermediate potential

Km

Kanamycin

LP

Low potential

MGDG

Monogalactosyldiacylglycerol

PG

Phosphatidylglycerol

PQ

Plastoquinone

PSI

Photosystem I

PSII

Photosystem II

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SQDG

Sulfoquinovosyldiacylglycerol

Notes

Acknowledgements

This work was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science (16J10129), and by CREST, Japan Science and Technology Agency.

Supplementary material

11120_2018_555_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 13 KB)
11120_2018_555_MOESM2_ESM.tif (6 mb)
Supplementary Fig. 1 BN-PAGE/SDS-PAGE analyses of PSII complexes purified from cells of the control strain and mutants. PSII complexes corresponding to 3 μg Chl were analyzed by BN-PAGE in the first dimension, and then by SDS-PAGE in the second dimension. Proteins in the gels of SDS-PAGE were visualized with silver staining. (TIF 6142 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Kaichiro Endo
    • 1
  • Koichi Kobayashi
    • 1
  • Hsing-Ting Wang
    • 2
  • Hsiu-An Chu
    • 2
  • Jian-Ren Shen
    • 3
  • Hajime Wada
    • 1
    Email author
  1. 1.Department of Life Sciences, Graduate School of Arts and SciencesThe University of TokyoTokyoJapan
  2. 2.Institute of Plant and Microbial BiologyAcademia SinicaTaipeiTaiwan, Republic of China
  3. 3.Research Institute for Interdisciplinary Science, Graduate School of Natural Science and TechnologyOkayama UniversityOkayamaJapan

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