Department of Cellular & Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
Proteomics. 2010 Aug;10(16):3045-50. doi: 10.1002/pmic.201000162.
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 approximately 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.
从纯化的细胞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物成员。当应用于纯化的核仁时，经典的核蛋白提取方法仅低效且选择性地释放约 50%的蛋白质。在这里，我们提出了一种可以提取高达 90%核仁蛋白的方法，并将其应用于定量相互作用组学方法中，以鉴定一种哺乳动物蛋白磷酸酶的核仁相互作用伙伴。
