Involvement of active oxygen species in degradation of the D1 protein under strong illumination in isolated subcomplexes of photosystem II.
The effects of strong illumination on the proteins in photosystem II (PSII) were investigated using three different isolated subcomplexes of PSII, namely, the PSII complex depleted of major light-harvesting proteins, the core complex, and the reaction center complex. Under illumination, not only the D1 protein of the reaction center but also other intrinsic proteins sustained some damage in all three subcomplexes: Coomassie blue-stained bands after polyacrylamide gel electrophoresis were smeared, and their migration distances on the gel were reduced with increasing duration of illumination. Such damage occurred first in the D1 and D2 proteins and subsequently in the 43- and 47-kDa proteins of the core antenna and the subunit of cytochrome b559. Immunoblot analysis using an antibody specific to the D1 protein showed that the D1 protein was degraded to major fragments of about 23 and 16 kDa during illumination. The smearing and changes in mobility of protein bands, as well as the fragmentation of the D1 protein, were greatly suppressed by scavengers of active oxygen species. From the effectiveness of scavengers, it appeared that superoxide anions participate in the protein damage in the PSII complex, hydrogen peroxide in the PSII and core complexes, and singlet oxygen, hydroxyl, and alkoxyl radicals in all three subcomplexes. We also found that fragments of the D1 protein of 23 and 16 kDa were formed even when PSII complexes that had been completely solubilized with sodium dodecyl sulfate were illuminated. This fragmentation was also suppressed by active oxygen scavengers.(ABSTRACT TRUNCATED AT 250 WORDS)
Miyao
M
M
Laboratory of Photosynthesis, National Institute of Agrobiological Resources (NIAR), Tsukuba, Japan.
D001426:Bacterial Proteins / Q000378:metabolism* / Q000528:radiation effects; D002374:Catalase / Q000378:metabolism; D016166:Free Radical Scavengers; D015151:Immunoblotting; D008027:Light / Q000009:adverse effects*; D012275:Oryza / Q000378:metabolism*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism* / Q000528:radiation effects; D045332:Photosystem II Protein Complex; D010940:Plant Proteins / Q000378:metabolism* / Q000528:radiation effects; D017382:Reactive Oxygen Species; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000378:metabolism
D001426:Bacterial Proteins; D016166:Free Radical Scavengers; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D017382:Reactive Oxygen Species; D015842:Serine Proteinase Inhibitors; D002374:Catalase; D013482:Superoxide Dismutase