Efficient extraction of nucleolar proteins for interactome analyses
Efficient extraction of nucleolar proteins for interactome analyses
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DOI：
10.1002/pmic.201000162
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发表时间：
2010-08-01
期刊：
影响因子：
3.4
通讯作者：
Trinkle-Mulcahy, Laura
中科院分区：
文献类型：
--
作者：
Chamousset, Delphine;Mamane, Sarah;Trinkle-Mulcahy, Laura
从纯化细胞器中高效提取蛋白质对于体外分析至关重要，包括通过基于亲和纯化的定量蛋白质组学方法鉴定蛋白质复合体成员。当应用于纯化的核仁时，经典的核蛋白提取方法效率低下，并且选择性地释放了大约50%的蛋白质。在这里，我们提出了一种可以提取高达90%的核仁蛋白的方法，并将其应用于定量相互作用方法中，以确定哺乳动物蛋白磷酸酶的核仁相互作用伙伴。
The efficient extraction of proteins from purified cellular organelles is critical for in vitro analyses, including identification of protein complex members by affinity purification-based quantitative proteomic approaches. When applied to purified nucleoli, classic nuclear protein extraction methods inefficiently and selectively release only similar to 50% of proteins. Here, we present a method that can extract up to 90% of nucleolar proteins, and apply it in a quantitative interactomic approach to identify nucleolar interaction partners for a mammalian protein phosphatase.