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基于iTRAQ的海洋硅藻三角褐指藻中与硅响应相关的代谢机制的蛋白质组学分析。

iTRAQ-based proteomic analysis of the metabolism mechanism associated with silicon response in the marine diatom Thalassiosira pseudonana.

作者信息

Du Chao, Liang Jun-Rong, Chen Dan-Dan, Xu Bin, Zhuo Wen-Hao, Gao Ya-Hui, Chen Chang-Ping, Bowler Chris, Zhang Wen

机构信息

School of Life Sciences, Xiamen University , Xiamen 361005, China.

出版信息

J Proteome Res. 2014 Feb 7;13(2):720-34. doi: 10.1021/pr400803w. Epub 2014 Jan 8.

Abstract

Silicon is a critical element for diatom growth; however our understanding of the molecular mechanisms involved in intracellular silicon responses are limited. In this study, an iTRAQ-LC-MS/MS quantitative proteomic approach was coupled with an established synchrony technique to reveal the global metabolic silicon-response in the model diatom Thalassiosira pseudonana subject to silicon starvation and readdition. Four samples, which corresponded to the time of silicon starvation, girdle band synthesis, valve formation, and right after daughter cell separation (0, 1, 5, 7 h), were collected for the proteomic analysis. The results indicated that a total of 1,831 proteins, representing 16% of the predicted proteins encoded by the T. pseudonana genome, could be identified. Of the identified proteins, 165 were defined as being differentially expressed proteins, and these proteins could be linked to multiple biochemical pathways. In particular, a number of proteins related to silicon transport, cell wall synthesis, and cell-cycle progress could be identified. In addition, other proteins that are potentially involved in amino acid synthesis, protein metabolism, and energy generation may have roles in the cellular response to silicon. Our findings provide a range of valuable information that will be of use for further studies of this important physiological response that is unique to diatoms.

摘要

硅是硅藻生长的关键元素;然而,我们对细胞内硅响应所涉及的分子机制的了解有限。在本研究中,一种iTRAQ-LC-MS/MS定量蛋白质组学方法与一种已建立的同步技术相结合,以揭示模式硅藻假微型海链藻在硅饥饿和重新添加硅的情况下的整体代谢性硅响应。收集了四个对应于硅饥饿、环带合成、瓣膜形成以及子细胞分离后即刻(0、1、5、7小时)的样本用于蛋白质组学分析。结果表明,总共可以鉴定出1831种蛋白质,占假微型海链藻基因组预测编码蛋白质的16%。在鉴定出的蛋白质中,165种被定义为差异表达蛋白质,这些蛋白质可与多种生化途径相关联。特别地,可以鉴定出一些与硅运输、细胞壁合成和细胞周期进程相关的蛋白质。此外,其他可能参与氨基酸合成、蛋白质代谢和能量产生的蛋白质可能在细胞对硅的响应中发挥作用。我们的研究结果提供了一系列有价值的信息,将有助于进一步研究硅藻特有的这一重要生理响应。

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