Specific degradation of the D1 protein of photosystem II by treatment with hydrogen peroxide in darkness: implications for the mechanism of degradation of the D1 protein under illumination.
The D1 protein of the photosystem II (PSII) reaction center has a rapid turnover and is specifically degraded under illumination in vivo. When isolated PSII membranes were treated in darkness with 10 mM hydrogen peroxide (H2O2), an active form of oxygen that is generated at the acceptor side of PSII under illumination, proteins of the PSII reaction center were specifically damaged in almost the same way as observed under illumination with strong light. The D1 protein and, to a lesser extent, the D2 protein were degraded to specific fragments, and cross-linked products (the covalently linked adduct of the D1 protein and the alpha subunit of cytochrome b559 and the heterodimer of the D1 and D2 proteins) were generated concomitantly. The site of cleavage of the D1 protein that gave rise to a major fragment of 22 kDa was located in the loop that connects membrane-spanning helixes IV and V. Treatment with H2O2 caused the same damage to proteins in isolated thylakoids and in core complexes that contained the non-heme iron at the acceptor side, but not in isolated reaction centers depleted of the iron. From these observations and the effects of reagents that are known to interact with the non-heme iron, it is suggested that the damage to proteins is caused by oxygen radicals generated by the non-heme iron in the Fe(II) state in a reaction with H2O2. It is proposed, moreover, that a similar mechanism is operative during the selective and specific degradation of the D1 protein under illumination.
Miyao
M
M
Laboratory of Photosynthesis, National Institute of Agrobiological Resources (NIAR), Tsukuba, Japan.
Ikeuchi
M
M
Yamamoto
N
N
Ono
T
T
D002736:Chloroplasts / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D003624:Darkness; D005609:Free Radicals; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000187:drug effects / Q000528:radiation effects; D008027:Light; D012275:Oryza; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000378:metabolism; D010788:Photosynthesis / Q000502:physiology*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D045332:Photosystem II Protein Complex
D005609:Free Radicals; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D006861:Hydrogen Peroxide