MALDI-TOF MS to monitor the kinetics of phospholipase A2-digestion of oxidized phospholipids.
MALDI-TOF MS to monitor the kinetics of phospholipase A2-digestion of oxidized phospholipids.
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DOI：
10.1016/j.ymeth.2015.12.013
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发表时间：
2016-07
期刊：
影响因子：
4.8
通讯作者：
J. Schröter;R. Süss;J. Schiller
中科院分区：
文献类型：
--
作者：
J. Schröter;R. Süss;J. Schiller
游离脂肪酸（FFA）通过磷脂酶A2（PLA2）释放，该酶在磷脂（PL）的n-2位置切割脂肪酸残基。在炎症性疾病中，活性氧（如HOCl）导致氧化修饰PL的形成（如氯醇生成）。目前还不清楚PL的氧化在多大程度上影响PLA2对它们的消化率。此外，对不饱和脂肪酸酰基残基位置（sn-1与sn-2位置）和头基修饰（例如磷脂酰胆碱（PC）与磷脂酰乙醇胺（PE））的影响的研究也缺乏。因此，本研究的目的是利用基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱法对这些方面进行研究，以阐明PL/溶血磷脂（LPL）比率作为pla2消化率的测量。我们将证明，HOCl对PL的氧化修饰对pla2的消化率有相当大的影响，即不饱和脂肪酸酰基残基的氧化导致PC和PE的消化率降低。此外，MALDI MS是一种方便可靠的研究相关变化的工具。
Free fatty acids (FFA) are released through phospholipase A2(PLA2), which cleaves the fatty acyl residue at thesn-2 position of phospholipids (PL). During inflammatory diseases, reactive oxygen species (such as HOCl) lead to the formation of oxidatively modified PL (e.g., chlorohydrin generation). It is still widely unknown to which extent the oxidation of PL influences their digestibility by PLA2. Additionally, investigations on the impact of the position of the unsaturated fatty acyl residue (sn-1 versussn-2 position) and modifications of the headgroup (for instance phosphatidylcholine (PC) versus phosphatidylethanolamine (PE)) are also lacking. Therefore, the aim of this study is the investigation of these aspects using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to elucidate the PL/lysophospholipid (LPL) ratios as measures of the PLA2digestibility.We will show that oxidative modifications of PL by HOCl have a considerable impact on the PLA2digestibility, i.e., oxidation of the unsaturated fatty acyl residues leads to a reduced digestibility of both PC and PE. Besides, it will be shown that MALDI MS is a convenient and reliable tool to investigate the related changes.