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莱茵衣藻细胞周期中脂质和代谢物的动态变化。

Dynamics of lipids and metabolites during the cell cycle of Chlamydomonas reinhardtii.

机构信息

Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany.

Brazilian Bioethanol Science and Technology Laboratory/CNPEM, Rua Giuseppe Máximo Scolfano 10000, 13083-970, Campinas, Brazil.

出版信息

Plant J. 2017 Oct;92(2):331-343. doi: 10.1111/tpj.13642. Epub 2017 Sep 8.

Abstract

Metabolites and lipids are the final products of enzymatic processes, distinguishing the different cellular functions and activities of single cells or whole tissues. Understanding these cellular functions within a well-established model system requires a systemic collection of molecular and physiological information. In the current report, the green alga Chlamydomonas reinhardtii was selected to establish a comprehensive workflow for the detailed multi-omics analysis of a synchronously growing cell culture system. After implementation and benchmarking of the synchronous cell culture, a two-phase extraction method was adopted for the analysis of proteins, lipids, metabolites and starch from a single sample aliquot of as little as 10-15 million Chlamydomonas cells. In a proof of concept study, primary metabolites and lipids were sampled throughout the diurnal cell cycle. The results of these time-resolved measurements showed that single compounds were not only coordinated with each other in different pathways, but that these complex metabolic signatures have the potential to be used as biomarkers of various cellular processes. Taken together, the developed workflow, including the synchronized growth of the photoautotrophic cell culture, in combination with comprehensive extraction methods and detailed metabolic phenotyping has the potential for use in in-depth analysis of complex cellular processes, providing essential information for the understanding of complex biological systems.

摘要

代谢物和脂质是酶促过程的最终产物,可区分单个细胞或整个组织的不同细胞功能和活动。要在成熟的模型系统中理解这些细胞功能,需要系统地收集分子和生理信息。在本报告中,选择绿藻莱茵衣藻来建立一个全面的工作流程,用于对同步生长的细胞培养系统进行详细的多组学分析。在实施和基准测试同步细胞培养后,采用两阶段提取方法从单个 10-1500 万个衣藻细胞的样品等分试样中分析蛋白质、脂质、代谢物和淀粉。在概念验证研究中,整个昼夜细胞周期中都采样了初级代谢物和脂质。这些时间分辨测量的结果表明,单个化合物不仅在不同途径中彼此协调,而且这些复杂的代谢特征有可能用作各种细胞过程的生物标志物。总之,所开发的工作流程包括光自养细胞培养的同步生长,结合全面的提取方法和详细的代谢表型分析,有可能用于深入分析复杂的细胞过程,为理解复杂的生物系统提供必要的信息。

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