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嗜热栖热菌在不同生长温度下差异表达的蛋白质及其在耐热适应中的作用。

Thermus thermophilus proteins that are differentially expressed in response to growth temperature and their implication in thermoadaptation.

机构信息

Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, The People's Republic of China.

出版信息

J Proteome Res. 2010 Feb 5;9(2):855-64. doi: 10.1021/pr900754y.

Abstract

As a kind of important extremophiles to realize the adaptation of life at high temperatures, thermophiles have attracted extensive studies. However, the pathways of thermophile proteins related to thermoadaptation remain to be addressed. Our study showed that there existed two types of protein profiles for the thermophile Thermus thermophilus wl in response to temperature change. One of them came from cultures growing below 65 degrees C, which was close to the optimal growth temperature, and another from cultures at or above 65 degrees C. These protein profiles were confirmed by Northern blots. On the basis of the proteomic and computational analyses, it was found that the thermophile proteins related to thermoadaptation might be involved in metabolic pathways as well as the stabilities and modifications of DNA and proteins. Interestingly, for the basic metabolism glycolysis, the phosphoglucomutase was up-regulated at below-optimum temperature, while the glyceraldehyde-3-phosphate dehydrogenase was up-regulated at above-optimum temperature, suggesting that different regulations might be used for basic metabolism at different temperatures. To characterize the proteins in response to high temperatures, superoxide dismutase (SOD), an important enzyme in organism to remove free radical produced in stress environment such as high temperature, was selected as a target protein for this investigation. SOD was inactivated to construct a SOD mutant. The results showed that the SOD protein was essential in thermoadaptation of T. thermophilus. Our study, therefore, presented the thermophile proteins required for thermoadaptation and their possible pathways in thermoadaptation.

摘要

作为实现高温生命适应的一种重要的极端生物,嗜热菌引起了广泛的研究。然而,与耐热适应相关的嗜热菌蛋白质途径仍有待解决。我们的研究表明,嗜热菌 Thermus thermophilus wl 存在两种应对温度变化的蛋白质谱类型。一种来自生长温度低于 65 摄氏度的培养物,接近最适生长温度,另一种来自生长温度在 65 摄氏度或以上的培养物。这些蛋白质谱通过 Northern blot 得到了证实。基于蛋白质组学和计算分析,发现与耐热适应相关的嗜热菌蛋白质可能涉及代谢途径以及 DNA 和蛋白质的稳定性和修饰。有趣的是,对于基本代谢途径糖酵解,磷酸葡糖变位酶在低于最适温度时上调,而甘油醛-3-磷酸脱氢酶在高于最适温度时上调,表明在不同温度下可能使用不同的调控方式进行基本代谢。为了研究对高温有反应的蛋白质,超氧化物歧化酶(SOD)作为一种重要的酶,可清除高温等应激环境中产生的自由基,被选为该研究的目标蛋白。SOD 被失活以构建 SOD 突变体。结果表明,SOD 蛋白在 Thermus thermophilus 的耐热适应中是必需的。因此,我们的研究提出了耐热适应所需的嗜热菌蛋白质及其可能的耐热适应途径。

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