A targeted mass spectrometric analysis of phosphatidylinositol phosphate species
A targeted mass spectrometric analysis of phosphatidylinositol phosphate species
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DOI：
10.1194/jlr.d500010-jlr200
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发表时间：
2005-08-01
影响因子：
6.5
通讯作者：
Brown, HA
中科院分区：
文献类型：
--
作者：
Milne, SB;Ivanova, PT;Brown, HA
一种新的质谱脂质分析方法的开发允许识别包括脂肪酰基组合物的细胞磷脂酰肌醇单磷酸/磷脂酰肌醇二磷酸/磷脂酰肌醇三磷酸（PIP/PIP 2/PIP 3）种类。使用电喷雾电离质谱，我们能够从RAW 264.7或原代鼠巨噬细胞提取物中解析和鉴定28种PIP和PIP 2化合物以及8种PIP 3化合物。激动剂刺激细胞后PIP谱的分析揭示了差异PIP 3种类的产生，并使我们能够提出一种新的方法来调节和特异性通过PIP 3的信号传导。这是首次报道的完整的，细胞PIP 3的质谱分析鉴定。分析信号脂质的脂肪酰基链组成的能力启动了新的场地调查的过程中，特定的聚磷酸肌醇物种介导。
The development of a new mass spectrometric lipid profiling methodology permits the identification of cellular phosphatidylinositol monophosphate/phosphatidylinositol bisphosphate/phosphatidylinositol trisphosphate (PIP/ PIP2/PIP3) species that includes the fatty acyl composition. Using electrospray ionization mass spectrometry, we were able to resolve and identify 28 PIP and PIP2 compounds as well as 8 PIP3 compounds from RAW 264.7 or primary murine macrophage cell extracts. Analysis of PIP profiles after agonist stimulation of cells revealed the generation of differential PIP3 species and permitted us to propose a novel means for regulation and specificity in signaling through PIP3. This is the first reported identification of intact, cellular PIP3 by mass spectral analysis. The ability to analyze the fatty acyl chain composition of signaling lipids initiates new venues for investigation of the processes by which specific polyphosphoinositide species mediate.