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Author:
Date:
2013-08-20
[Abstract] The lipid and protein interactions are an integral and important part of many cellular signaling pathways. The understanding of the selective and specific interaction of the given lipid molecule with the target protein is required for studying cellular signaling. In this assay, different lipids are spotted onto a nitrocellulose membrane to which they attach. Then the membrane is incubated with a lipid binding protein possessing an epitope tag. The protein binds to the lipid which is detected by immunoblotting with an antibody recognizing the epitope tag (see Figure 1). PTEN is an important tumor suppressor which functions as both protein and lipid phosphatase. The primary physiological substrate of PTEN is signaling lipid PtdIns (3, 4, 5) P3, by dephosphrylating PtdIns (3, 4, 5) P3 to ...
[摘要] 脂质和蛋白质相互作用是许多细胞信号传导途径的整体和重要部分。研究细胞信号传导需要理解给定脂质分子与靶蛋白的选择性和特异性相互作用。在该测定中，将不同的脂质点在它们附着的硝酸纤维素膜上。然后将膜与具有表位标签的脂质结合蛋白一起温育。蛋白质与脂质结合，其通过用识别表位标签的抗体的免疫印迹检测（参见图1）。 PTEN是一种重要的肿瘤抑制剂，其作为蛋白质和脂质磷酸酶起作用。 PTEN的主要生理学底物是信号传导脂质PtdIns（3,4,5）P3，通过将PtdIns（3,4,5）P3去磷酸化为PtdIns（4,5）P2PTEN负调节PI3K信号传导并介导其肿瘤抑制功能通过灭活下游致癌AKT介导的信号。进行PTEN脂质结合测定以研究PTEN与不同脂质分子的特异性结合。
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