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iTRAQ 定量蛋白质组学分析揭示了节旋藻耐热性的潜在变化。

Quantitative proteomics analysis by iTRAQ revealed underlying changes in thermotolerance of Arthrospira platensis.

机构信息

College of Biological Sciences and Technology, Beijing Forestry University, No. 35 Qinghua east road, Haidian District, Beijing, China.

College of Biological Sciences and Technology, Beijing Forestry University, No. 35 Qinghua east road, Haidian District, Beijing, China.

出版信息

J Proteomics. 2017 Aug 8;165:119-131. doi: 10.1016/j.jprot.2017.06.015. Epub 2017 Jun 20.

Abstract

UNLABELLED

Growth temperature is a critical factor that affects cultivation of Arthrospira platensis which is a type of cyanobacterium widely known as Spirulina that has significant commercial value. To investigate the molecular mechanism underlying the thermotolerance of Spirulina, differential protein expression profiling was carried out using iTRAQ-based quantitative proteomic analysis. This study only analyzed changes in thylakoids. Among the 2085 proteins quantified, 43 differentially expressed proteins were selected based on the fold change cutoff scores of ≥2 or ≤0.5 for up-regulation or down-regulation, respectively. An analysis of these 43 proteins found that 23% of them are photosynthetic system proteins which include photosynthetic enzymes and pigment proteins. The dynamic change of these proteins indicates that photosynthetic system functions were profoundly affected under heat stress and the light-dependent reactions were probably the most sensitive to temperature changes. Meanwhile, to cope with the low energy production due to impaired photosynthesis there was a considerable down-shift in protein synthesis which is a very energy demanding process. The impaired photosynthesis led to low energy generation that was compensated by a down-shift in translation (the most energy-demanding process) and an up-shift of glycolysis. The reduction of many ribosome proteins may lead to a loss in translation efficiency; therefore, Spirulina may adopted a different mechanism to increase translational elongation under heat stress to compensate for this loss, such as elevate L7/L12 proteins. Changes were also found in the classical heat shock proteins, the ROS scavenging system, DNA-binding proteins, and some membrane proteins. In conclusion, this research demonstrate that heat stress induces profound changes in cellular physiology and shed light on the mechanism of the heat stress response and thermotolerance of Arthrospira platensis.

BIOLOGICAL SIGNIFICANCE

Arthrospira platensis, widely known as Spirulina, is a type of cyanobacteria which is cultivated at large scale for it great commercial value. It has become a consensus that continually increasing temperature due to global warming is bringing serious threat to agriculture, including the Spirulina cultivation. High temperature not only limits biomass accumulation by Spirulina, but also changes the composition of nutrition. Therefore there is a greater need than ever before to understand how Spirulina tolerates and cope with high temperature. In this study, we for the first time applied the iTRAQ-based quantitative proteomic technology to investigate the thermotolerance of Spirulina. Our results showed that many biological processes were altered by heat stress. Most significantly, we found that heat stress harmed the photosynthesis ability and caused low energy production, and to deal with this situation, energy demanding processes like protein synthesis were down-shifted and the alternative energy metabolism process glycolysis was up-shifted. Our results also show other important proteins, like the classical heat shock proteins and some antioxidant proteins, are also increased. Thus our study sheds light to our understanding of the mechanism underlying the thermotolerance of Spirulina.

摘要

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生长温度是影响广泛称为螺旋藻的蓝藻培养的关键因素,螺旋藻具有重要的商业价值。为了研究螺旋藻耐热性的分子机制,我们使用 iTRAQ 定量蛋白质组学分析进行了差异蛋白质表达谱分析。本研究仅分析了类囊体的变化。在定量的 2085 种蛋白质中,根据上调或下调分别为≥2 或≤0.5 的折叠变化截止分数,选择了 43 种差异表达的蛋白质。对这 43 种蛋白质的分析表明,其中 23%是光合作用系统蛋白,包括光合作用酶和色素蛋白。这些蛋白质的动态变化表明,光合作用系统功能在热应激下受到严重影响,光依赖反应可能对温度变化最敏感。同时,为了应对光合作用受损导致的能量产生减少,蛋白质合成有相当大的下调,这是一个非常耗能的过程。受损的光合作用导致能量产生减少,通过翻译的下调(最耗能的过程)和糖酵解的上调来补偿。许多核糖体蛋白的减少可能导致翻译效率降低;因此,螺旋藻可能采用不同的机制在热应激下增加翻译延伸,以弥补这种损失,例如上调 L7/L12 蛋白。经典热休克蛋白、ROS 清除系统、DNA 结合蛋白和一些膜蛋白也发生了变化。总之,本研究表明,热应激会引起细胞生理学的深刻变化,并阐明了 Arthrospira platensis 耐热性的热应激反应机制。

生物学意义

广泛称为螺旋藻的 Arthrospira platensis 是一种蓝藻,因其巨大的商业价值而被大规模种植。人们已经达成共识,由于全球变暖导致的气温持续升高,正在给农业带来严重威胁,包括螺旋藻的种植。高温不仅限制了螺旋藻的生物量积累,还改变了营养成分的组成。因此,比以往任何时候都更需要了解螺旋藻如何耐受和应对高温。在这项研究中,我们首次应用 iTRAQ 定量蛋白质组学技术来研究螺旋藻的耐热性。我们的结果表明,许多生物过程受到热应激的影响。最重要的是,我们发现热应激损害了光合作用能力并导致能量产生减少,为了应对这种情况,像蛋白质合成这样耗能的过程被下调,而替代的能量代谢过程糖酵解被上调。我们的结果还显示了其他重要的蛋白质,如经典的热休克蛋白和一些抗氧化蛋白,也增加了。因此,我们的研究结果阐明了螺旋藻耐热性的机制。

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