Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis.
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 micromol 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 degrees 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.
Department of Biology, Faculty of Science, Tokyo University of Science, Kagurazaka 1-3, Shinjuku-ku, Tokyo 162-8601, Japan.
Ryo
R
Nagao
Tatsuya
T
Tomo
Eri
E
Noguchi
Saori
S
Nakajima
Takehiro
T
Suzuki
Akinori
A
Okumura
Yasuhiro
Y
Kashino
Mamoru
M
Mimuro
Masahiko
M
Ikeuchi
Isao
I
Enami
D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D017377:Diatoms / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D045342:Light-Harvesting Protein Complexes / Q000302:isolation & purification / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D045332:Photosystem II Protein Complex / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D010971:Plastoquinone / Q000378:metabolism
D045342:Light-Harvesting Protein Complexes; D045332:Photosystem II Protein Complex; C088752:fucoxanthin-, chlorophyll a-c-containing protein; D002734:Chlorophyll; D010971:Plastoquinone; D010100:Oxygen; D000077194:Chlorophyll A