Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis

Ryo Nagao, Tatsuya Tomo, Eri Noguchi, Saori Nakajima, Takehiro Suzuki, Akinori Okumura, Yasuhiro Kashino, Mamoru Mimuro, Masahiko Ikeuchi, Isao Enami

Research output: Contribution to journalArticle

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Abstract

Oxygen-evolving Photosystem II particles (crude PSII) retaining a high oxygen-evolving activity have been prepared from a marine centric diatom, Chaetoceros gracilis (Nagao et al., 2007). The crude PSII, however, contained a large amount of fucoxanthin chlorophyll a/c-binding proteins (FCP). In this study, a purified PSII complex which was deprived of major components of FCP was isolated by one step of anion exchange chromatography from the crude PSII treated with Triton X-100. The purified PSII was still associated with the five extrinsic proteins of PsbO, PsbQ', PsbV, Psb31 and PsbU, and showed a high oxygen-evolving activity of 2135 μmol O2 (mg Chl a)- 1 h- 1 in the presence of phenyl-p-benzoquinone which was virtually independent of the addition of CaCl2. This activity is more than 2.5-fold higher than the activity of the crude PSII. The activity was completely inhibited by 3-(3,4)-dichlorophenyl-(1,1)-dimethylurea (DCMU). The purified PSII contained 42 molecules of Chl a, 2 molecules of diadinoxanthin and 2 molecules of Chl c on the basis of two molecules of pheophytin a, and showed typical absorption and fluorescence spectra similar to those of purified PSIIs from the other organisms. In this study, we also found that the crude PSII was significantly labile, as a significant inactivation of oxygen evolution, chlorophyll bleaching and degradation of PSII subunits were observed during incubation at 25 °C in the dark. In contrast, these inactivation, bleaching and degradation were scarcely detected in the purified PSII. Thus, we succeeded for the first time in preparation of a stable PSII from diatom cells.

Original languageEnglish
Pages (from-to)160-166
Number of pages7
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1797
Issue number2
DOIs
Publication statusPublished - Feb 2010
Externally publishedYes

Fingerprint

Diatoms
Photosystem II Protein Complex
Purification
Diuron
Oxygen
Molecules
Bleaching
Chlorophyll Binding Proteins
Degradation
Octoxynol
Chlorophyll
Chromatography
Anions
Fluorescence
Proteins

Keywords

  • Chaetoceros gracilis
  • Diatom
  • Extrinsic protein
  • Oxygen evolution
  • Photosystem II

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis. / Nagao, Ryo; Tomo, Tatsuya; Noguchi, Eri; Nakajima, Saori; Suzuki, Takehiro; Okumura, Akinori; Kashino, Yasuhiro; Mimuro, Mamoru; Ikeuchi, Masahiko; Enami, Isao.

In: Biochimica et Biophysica Acta - Bioenergetics, Vol. 1797, No. 2, 02.2010, p. 160-166.

Research output: Contribution to journalArticle

Nagao, Ryo ; Tomo, Tatsuya ; Noguchi, Eri ; Nakajima, Saori ; Suzuki, Takehiro ; Okumura, Akinori ; Kashino, Yasuhiro ; Mimuro, Mamoru ; Ikeuchi, Masahiko ; Enami, Isao. / Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis. In: Biochimica et Biophysica Acta - Bioenergetics. 2010 ; Vol. 1797, No. 2. pp. 160-166.
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AU - Suzuki, Takehiro

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AU - Kashino, Yasuhiro

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