Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Biochemistry (Mosc). 2011 Apr;76(4):427-37. doi: 10.1134/s0006297911040055.
The rate of PSI mediated cyclic electron transport was studied in wild type and mutant cells of Synechocystis sp. PCC 6803 deficient in NDH-1 (M55) or succinate dehydrogenase (SDH(-)) that are responsible for the dark reduction of the plastoquinone pool. Kinetics of P700 photooxidation and P700(+) dark reduction in the presence of 5·10(-5) M 3-(3,4-dichlorophenyl)-1,1-dimethylurea have been registered as light induced absorbance changes at 810 nm resulting from illumination of cells with 730-nm actinic light for 1 sec. It is shown that in the absence of dehydrogenases the rate of dark reduction of P700(+) in both mutants did not decrease but even increased in NDH-1-less mutant cells as compared with the rate in wild type cells. Dibromothymoquinone drastically reduced the rate of P700(+) dark reduction both in wild type and in mutant cells. Thus, the cyclic electron transfer from ferredoxin through the plastoquinone pool to P700(+), which is independent from dehydrogenases, takes place in all the types of cells. Preillumination of cells of wild type and both mutants for 30 min or anaerobic conditions resulted in delay of P700 photooxidation and acceleration of P700(+) dark reduction, while the level of photosynthesis and respiration terminal acceptors (NAD(P)(+) and oxygen) decreased. It appears that the rate of P700 photooxidation and P700(+) dark reduction in cyclic electron transport in Synechocystis wild type and mutant cells is determined by the level of NADP+ and oxygen in stroma. A possible approach to evaluation of the levels of these acceptors in vivo is proposed, based on kinetic curve parameters of P700 photoconversions induced by 730-nm light with 1-sec duration.
研究了野生型和突变型 Synechocystis sp. PCC 6803 细胞中 PSI 介导的环式电子传递速率,这些细胞缺乏负责质醌池暗还原的 NDH-1(M55)或琥珀酸脱氢酶(SDH(-))。在存在 5·10(-5) M 3-(3,4-二氯苯基)-1,1-二甲基脲的情况下,记录了 P700 光氧化和 P700(+)暗还原的动力学,方法是将细胞用 730nm 光照射 1 秒,记录 810nm 处的光诱导吸收变化。结果表明,在没有脱氢酶的情况下,与野生型细胞相比,NDH-1 缺失突变体细胞中 P700(+)的暗还原速率没有降低,甚至增加。二溴噻吩醌(Dibromothymoquinone)大大降低了野生型和突变型细胞中 P700(+)的暗还原速率。因此,来自铁氧还蛋白的电子通过质醌池传递到 P700(+)的环式电子传递,这种传递不依赖于脱氢酶,发生在所有类型的细胞中。细胞预照 30 分钟或厌氧条件会导致 P700 光氧化延迟和 P700(+)暗还原加速,而光合作用和呼吸作用末端受体(NAD(P)(+)和氧气)的水平降低。看来,Synechocystis 野生型和突变型细胞中环式电子传递中 P700 光氧化和 P700(+)暗还原的速率取决于基质中 NADP+和氧气的水平。提出了一种基于 730nm 光持续 1 秒诱导的 P700 光转化动力学曲线参数来评估体内这些受体水平的方法。