Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)
Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)
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DOI：
10.3390/metabo2010057
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发表时间：
2012-03-01
期刊：
影响因子：
4.1
通讯作者：
Bruegger, Britta
中科院分区：
文献类型：
--
作者：
Haag, Mathias;Schmidt, Angelika;Bruegger, Britta
磷脂（PIPs）和二酰基甘油（DAG）等脂质是参与细胞过程（如T细胞受体（TCR）介导的信号转导）的重要信号中间体。本文报道了粗脂提取物中PIP、PIP2和DAG的鉴定和定量。利用pip和dag的不同提取特性，我们可以有效地从一个样品中回收这两类脂质。采用纳米电喷雾电离串联质谱（nano-ESI MS/MS）对内源性信号分子进行快速分析，采用脂类特异性中性损失和多重前体离子扫描对其进行鉴定和定量。在TCR刺激前后，对原代人T细胞中DAG、PIP和PIP2分子种类的分析导致DAG水平增加两倍，并在刺激细胞中向1-硬脂酰-2-花生四烯酰基-DAG转变。PIP2水平略有下降，而PIP水平保持不变。
Lipids, such as phosphoinositides (PIPs) and diacylglycerol (DAG), are important signaling intermediates involved in cellular processes such as T cell receptor (TCR)-mediated signal transduction. Here we report identification and quantification of PIP, PIP2 and DAG from crude lipid extracts. Capitalizing on the different extraction properties of PIPs and DAGs allowed us to efficiently recover both lipid classes from one sample. Rapid analysis of endogenous signaling molecules was performed by nano-electrospray ionization tandem mass spectrometry (nano-ESI MS/MS), employing lipid class-specific neutral loss and multiple precursor ion scanning for their identification and quantification. Profiling of DAG, PIP and PIP2 molecular species in primary human T cells before and after TCR stimulation resulted in a two-fold increase in DAG levels with a shift towards 1-stearoyl-2-arachidonoyl-DAG in stimulated cells. PIP2 levels were slightly reduced, while PIP levels remained unchanged.