Photosynthesis Research

, Volume 126, Issue 2–3, pp 385–397 | Cite as

Site-directed mutagenesis of amino acid residues of D1 protein interacting with phosphatidylglycerol affects the function of plastoquinone QB in photosystem II

  • Kaichiro Endo
  • Naoki Mizusawa
  • Jian-Ren Shen
  • Masato Yamada
  • Tatsuya Tomo
  • Hirohisa Komatsu
  • Masami Kobayashi
  • Koichi Kobayashi
  • Hajime Wada
Regular Paper

Abstract

Recent X-ray crystallographic analysis of photosystem (PS) II at 1.9-Å resolution identified 20 lipid molecules in the complex, five of which are phosphatidylglycerol (PG). In this study, we mutagenized amino acid residues S232 and N234 of D1, which interact with two of the PG molecules (PG664 and PG694), by site-directed mutagenesis in Synechocystis sp. PCC 6803 to investigate the role of the interaction in PSII. The serine and asparagine residues at positions 232 and 234 from the N-terminus were mutagenized to alanine and aspartic acid, respectively, and a mutant carrying both amino acid substitutions was also produced. Although the obtained mutants, S232A, N234D, and S232AN234D, exhibited normal growth, they showed decreased photosynthetic activities and slower electron transport from QA to QB than the control strain. Thermoluminescence analysis suggested that this slower electron transfer in the mutants was caused by more negative redox potential of QB, but not in those of QA and S2. In addition, the levels of extrinsic proteins, PsbV and PsbU, were decreased in PSII monomer purified from the S232AN234D mutant, while that of Psb28 was increased. In the S232AN234D mutant, the content of PG in PSII was slightly decreased, whereas that of monogalactosyldiacylglycerol was increased compared with the control strain. These results suggest that the interactions of S232 and N234 with PG664 and PG694 are important to maintain the function of QB and to stabilize the binding of extrinsic proteins to PSII.

Keywords

D1 Phosphatidylglycerol Photosystem II Site-directed mutagenesis Synechocystis sp. PCC 6803 

Abbreviation

BQ

p-Benzoquinone

Chl

Chlorophyll

Cm

Chloramphenicol

CmR

Chloramphenicol-resistant gene cassette

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

Fecy

Potassium ferricyanide

Km

Kanamycin

KmR

Kanamycin-resistant gene cassette

MGDG

Monogalactosyldiacylglycerol

PCR

Polymerase chain reaction

PG

Phosphatidylglycerol

PQ

Plastoquinone

PSI

Photosystem I

PSII

Photosystem II

SDS-PAGE

SDS-polyacrylamide gel electrophoresis

SQDG

Sulfoquinovosyldiacylglycerol

Spe/Str

Spectinomycin/streptomycin

Spe/StrR

Spectinomycin/streptomycin-resistant gene cassette

Tet

Tetracycline

TetR

Tetracycline-resistant gene cassette

WT

Wild-type

Supplementary material

11120_2015_150_MOESM1_ESM.docx (23.2 mb)
Supplementary material 1 (DOCX 23764 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Kaichiro Endo
    • 1
  • Naoki Mizusawa
    • 2
    • 3
  • Jian-Ren Shen
    • 4
  • Masato Yamada
    • 5
  • Tatsuya Tomo
    • 5
  • Hirohisa Komatsu
    • 6
  • Masami Kobayashi
    • 6
  • Koichi Kobayashi
    • 1
  • Hajime Wada
    • 1
  1. 1.Department of Life Sciences, Graduate School of Arts and SciencesThe University of TokyoTokyoJapan
  2. 2.Department of Frontier BioscienceHosei UniversityKoganeiJapan
  3. 3.Research Center for Micro-Nano TechnologyHosei UniversityKoganeiJapan
  4. 4.Photosynthesis Research Center, Graduate School of Natural Science and TechnologyOkayama UniversityOkayamaJapan
  5. 5.Faculty of ScienceTokyo University of ScienceTokyoJapan
  6. 6.Division of Materials Science, Faculty of Pure and Applied ScienceUniversity of TsukubaIbarakiJapan

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