Isolation of the Plant Cytosolic Fraction for Proteomic Analysis
Isolation of the Plant Cytosolic Fraction for Proteomic Analysis
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DOI：
10.1007/978-1-62703-631-3_31
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发表时间：
2014-01-01
期刊：
影响因子：
--
通讯作者：
Ito, Jun
中科院分区：
文献类型：
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作者：
Estavillo, Gonzalo M.;Verhertbruggen, Yves;Ito, Jun
细胞质是细胞中未被膜分隔的液体部分。它包含高度多样化的物质集合，是许多重要细胞过程的核心，从信号转导、代谢物产生和运输、蛋白质生物合成和降解到应激反应和防御。尽管其重要性，但仅对植物细胞质进行了少量蛋白质组学研究。这主要是由于从不含破坏的细胞器材料的植物材料中分离出相对纯净的样品存在困难。在本章中，我们概述了从拟南芥细胞培养物和幼苗中分离胞质部分的方法，并提供了评估质谱分析纯度的指导。
The cytosol is the fluid portion of the cell that is not partitioned by membranes. It contains a highly diverse collection of substances and is central to many essential cellular processes ranging from signal transduction, metabolite production and transport, protein biosynthesis and degradation to stress response and defense. Despite its importance, only a few proteomic studies have been performed on the plant cytosol. This is largely due to difficulties in isolating relatively pure samples from plant material free of disrupted organelle material. In this chapter we outline methods for isolating the cytosolic fraction from Arabidopsis cell cultures and seedlings and provide guidance on assessing purity for analysis by mass spectrometry.