Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.
Graduate School of Science, Kobe University, Kobe, Hyogo 657-8501, Japan.
J Proteomics. 2021 Aug 30;246:104306. doi: 10.1016/j.jprot.2021.104306. Epub 2021 Jun 19.
In cyanobacteria, it is known that the excitation ratios of photosystem (PS) I and PSII changes with the wavelength of irradiated light due to mobile phycobilisome (PBS) and spillover, affecting the photosynthetic ATP/NADPH synthesis ratio and metabolic flux state. However, the mechanisms by which these changes are controlled have not been well studied. In this study, we performed a targeted proteomic analysis of Synechocystis sp. PCC 6803 under different spectral light conditions to clarify the regulation mechanisms of mobile PBS, spillover and metabolisms under different light qualities at the protein level. The results showed an increase in the amount of proteins mainly involved in CO fixation under Red1 light conditions with a high specific growth rate, suggesting that the rate of intracellular metabolism is controlled by the rate of carbon uptake, not by changes in the amount of each enzyme. Correlation analysis between protein levels and PSI/PSII excitation ratios revealed that PsbQUY showed high correlations and significantly increased under Blue and Red2 light conditions, where the PSI/PSII excitation ratio was higher due to spillover. In the strains lacking the genes encoding these proteins, a decrease in the PSI/PSII excitation ratio was observed, suggesting that PsbQUY contribute to spillover occurrence. SIGNIFICANCE: In cyanobacteria, the photosynthetic apparatus's responses, such as state transition [mobile PBS and spillover], occur due to the intensity and wavelength of irradiated light, resulting in changes in photosynthetic electron transport and metabolic flux states. Previous studies have analyzed the response of Synechocystis sp. PCC 6803 to light intensity from various directions, but only spectroscopic analysis of the photosynthetic apparatus has been done on the response to changes in the wavelength of irradiated light. This study analyzed the response mechanisms of mobile PBS, spillover, photosynthetic, and metabolic systems in Synechocystis sp. PCC 6803 under six different spectral light conditions by a targeted proteomic analysis. As a result, many proteins were successfully quantified, and the metabolic enzymes and photosynthetic apparatus were analyzed using an integrated approach. Principal component and correlation analyses and volcano plots revealed that the PSII subunits PsbQ, PsbU, and PsbY have a strong correlation with the PSI/PSII excitation ratio and contribute to spillover occurrence. Thus, statistical analysis based on proteome data revealed that PsbQ, PsbU, and PsbY are involved in spillover, as revealed by spectroscopic analysis.
在蓝藻中,由于可移动的藻胆体(PBS)和溢出现象,光系统(PS)I 和 PSII 的激发比会随照射光的波长而变化,这会影响光合 ATP/NADPH 的合成比和代谢通量状态。然而,这些变化的控制机制尚未得到很好的研究。在这项研究中,我们对不同光谱光条件下的集胞藻 PCC 6803 进行了靶向蛋白质组学分析,以在蛋白质水平上阐明在不同光质下可移动 PBS、溢出现象和代谢的调控机制。结果表明,在高比生长率的红光 1 条件下,主要涉及 CO2 固定的蛋白质数量增加,这表明细胞内代谢的速率是由碳摄取的速率控制的,而不是由每种酶的数量变化控制的。蛋白质水平与 PSI/PSII 激发比的相关分析表明,在由于溢出现象而 PSI/PSII 激发比较高的蓝光和红光 2 条件下,PsbQUY 表现出高度相关性并显著增加。在缺乏编码这些蛋白质的基因的菌株中,观察到 PSI/PSII 激发比降低,表明 PsbQUY 有助于溢出现象的发生。意义:在蓝藻中,光合作用装置的响应(如状态转换[可移动 PBS 和溢出现象])是由于照射光的强度和波长引起的,这导致光合电子传递和代谢通量状态发生变化。先前的研究分析了集胞藻 PCC 6803 对来自不同方向的光强度的响应,但仅对光合作用装置对照射光波长变化的响应进行了分光分析。本研究通过靶向蛋白质组学分析,分析了集胞藻 PCC 6803 在六种不同光谱光条件下可移动 PBS、溢出现象、光合作用和代谢系统的响应机制。结果成功地定量了许多蛋白质,并使用集成方法分析了代谢酶和光合作用装置。主成分和相关性分析以及火山图表明,PSII 亚基 PsbQ、PsbU 和 PsbY 与 PSI/PSII 激发比具有很强的相关性,并且有助于溢出现象的发生。因此,基于蛋白质组数据的统计分析表明,分光分析表明 PsbQ、PsbU 和 PsbY 参与溢出现象。