Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University, Šlechtitelů 11, 783 71 Olomouc, Czech Republic.
Plant Cell Environ. 2012 Apr;35(4):806-18. doi: 10.1111/j.1365-3040.2011.02454.x. Epub 2011 Dec 13.
The cyanobacterial small CAB-like proteins (SCPs) are single-helix membrane proteins mostly associated with the photosystem II (PSII) complex that accumulate under stress conditions. Their function is still ambiguous although they are assumed to regulate chlorophyll (Chl) biosynthesis and/or to protect PSII against oxidative damage. In this study, the effect of SCPs on the PSII-specific light-induced damage and generation of singlet oxygen ((1)O(2)) was assessed in the strains of the cyanobacterium Synechocystis sp. PCC 6803 lacking PSI (PSI-less strain) or lacking PSI together with all SCPs (PSI-less/scpABCDE(-) strain). The light-induced oxidative modifications of the PSII D1 protein reflected by a mobility shift of the D1 protein and by generation of a D1-cytochrome b-559 adduct were more pronounced in the PSI-less/scpABCDE(-) strain. This increased protein oxidation correlated with a faster formation of (1)O(2) as detected by the green fluorescence of Singlet Oxygen Sensor Green assessed by a laser confocal scanning microscopy and by electron paramagnetic resonance spin-trapping technique using 2, 2, 6, 6-tetramethyl-4-piperidone (TEMPD) as a spin trap. In contrast, the formation of hydroxyl radicals was similar in both strains. Our results show that SCPs prevent (1)O(2) formation during PSII damage, most probably by the binding of free Chl released from the damaged PSII complexes.
蓝细菌小 CAB 样蛋白(SCPs)是与光系统 II(PSII)复合物主要相关的单螺旋膜蛋白,在应激条件下积累。尽管它们被认为可以调节叶绿素(Chl)生物合成和/或保护 PSII 免受氧化损伤,但它们的功能仍然不清楚。在这项研究中,评估了 SCPs 在缺乏 PSI(PSI-less 菌株)或缺乏 PSI 以及所有 SCPs(PSI-less/scpABCDE(-)菌株)的蓝细菌 Synechocystis sp. PCC 6803 菌株中对 PSII 特异性光诱导损伤和单线态氧((1)O(2))生成的影响。通过 D1 蛋白的迁移变化和 D1-细胞色素 b-559 加合物的生成反映的 PSII D1 蛋白的光诱导氧化修饰在 PSI-less/scpABCDE(-)菌株中更为明显。这种增加的蛋白质氧化与通过激光共聚焦扫描显微镜评估的 Singlet Oxygen Sensor Green 的绿色荧光检测到的更快的(1)O(2)形成相关,通过使用 2,2,6,6-四甲基-4-哌啶酮(TEMPD)作为自旋捕获剂的电子顺磁共振自旋捕获技术也可检测到(1)O(2)形成。相比之下,两种菌株中羟基自由基的形成相似。我们的结果表明,SCPs 可防止 PSII 损伤过程中(1)O(2)的形成,这很可能是通过结合从受损 PSII 复合物中释放的游离 Chl 实现的。