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Author:
Date:
2014-08-05
[Abstract] Diacylglycerol (DAG) is a bioactive lipid with diverse biological roles. DAG transiently accumulates in a membrane upon receipt of an appropriate stimulus that activates phospholipase C to cleave phospholipids. The resulting hydrolysis product DAG binds to proteins such as protein kinase C to initiate a variety of downstream cellular processes. DAG kinases attenuate such responses by converting DAG to phosphatidic acid.
This protocol describes an assay designed to quantify cellular DAG levels. The assay exploits the enzymatic conversion of DAG (sn-1,2-diacylglycerol) to phosphatidic acid (1,2-diacyl- sn-glycerol-3-phosphate) in conjunction with the incorporation of a radiolabeled phosphate group by DAG kinase (Figure 1). This assay was described in (Strijbis et ...
[摘要] 二酰基甘油（DAG）是具有不同生物学作用的生物活性脂质。 DAG在接受激活磷脂酶C以切割磷脂的适当刺激时在膜中瞬时累积。 所得水解产物DAG结合蛋白质例如蛋白激酶C以启动各种下游细胞过程。 DAG激酶通过将DAG转化为磷脂酸来减弱这种应答。该方案描述了设计用于定量细胞DAG水平的测定。 该测定利用DAG（1,2-二酰基甘油）到磷脂酸（1,2-二酰基-sn-甘油-3-磷酸）的酶促转化 结合DAG激酶掺入放射性标记的磷酸基团（图1）。 该测定在（Strijbis等人，2013）中描述。
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