Shui Guanghou, Guan Xue Li, Low Choon Pei, Chua Gek Huey, Goh Joyce Sze Yuin, Yang Hongyuan, Wenk Markus R
Yong Loo Lin School of Medicine, National University of Singapore, Department of Biochemistry, 8 Medical Drive, Block MD 7, Singapore.
Mol Biosyst. 2010 Jun;6(6):1008-17. doi: 10.1039/b913353d. Epub 2010 Feb 25.
Recent rapid growth of lipidomics is mainly attributed to technological advances in mass spectrometry. Development of soft ionization techniques, in combination with computational tools, has spurred subsequent development of various methods for lipid analysis. However, none of these existing approaches can cover major cellular lipids in a single run. Here we demonstrate that a single method of liquid chromatography coupled with mass spectrometry (LCMS) can be used for simultaneous profiling of major cellular lipids including glycerophospholipids (PLs), sphingolipids (SPLs), waxes, sterols (ST) and mono-, di- as well as triacylglycerides (MAG, DAG, TAG). We applied this approach to analyze these lipids in various organisms including Saccharomyces cerevisiae and Schizosaccharomyces pombe. While phospholipids and triacylglycerides of S. pombe mainly contain 18 : 1 fatty acyls, those of S. cerevisiae contain 16 : 1, 16 : 0 and 18 : 1 fatty acyls. S. cerevisiae and S. pombe contain distinct sphingolipid profiles. S. cerevisiae has abundant inositol phytoceramides (IPC), while S. pombe contains high levels of free phytoceramides as well as short chain phytoceramides (t18:1/20 : 0-B) and IPC (t18:1/20 : 0-B). In S. cerevisiae, our results demonstrated accumulation of ergosterol esters in tgl1Delta cells and accumulation of various TAG species in tgl3Delta cells, which are consistent with the function of the respective enzymes. Furthermore, we, for the first time, systematically characterized lipids in S. pombe and measured their dynamic changes in Deltaplh1Deltadga1 cells at different growth phases. We further discussed dynamic changes of phospholipids, sphingolipids and neutral lipids in the progress of programmed cell death in Deltaplh1Deltadga1 cells of S. pombe.
脂质组学最近的快速发展主要归因于质谱技术的进步。软电离技术与计算工具相结合，推动了各种脂质分析方法的后续发展。然而，这些现有方法中没有一种能在一次运行中涵盖主要的细胞脂质。在此，我们证明液相色谱-质谱联用（LCMS）的单一方法可用于同时分析包括甘油磷脂（PLs）、鞘脂（SPLs）、蜡、固醇（ST）以及单酰基甘油、二酰基甘油和三酰基甘油（MAG、DAG、TAG）在内的主要细胞脂质。我们应用此方法分析了包括酿酒酵母和粟酒裂殖酵母在内的各种生物体中的这些脂质。粟酒裂殖酵母的磷脂和三酰基甘油主要含有18 : 1脂肪酸酰基，而酿酒酵母的则含有16 : 1、16 : 0和18 : 1脂肪酸酰基。酿酒酵母和粟酒裂殖酵母具有不同的鞘脂谱。酿酒酵母富含肌醇植物神经酰胺（IPC），而粟酒裂殖酵母含有高水平的游离植物神经酰胺以及短链植物神经酰胺（t18:1/20 : 0-B）和IPC（t18:1/20 : 0-B）。在酿酒酵母中，我们的结果表明在tgl1Delta细胞中麦角固醇酯积累，在tgl3Delta细胞中各种TAG种类积累，这与相应酶的功能一致。此外，我们首次系统地表征了粟酒裂殖酵母中的脂质，并测量了Deltaplh1Deltadga1细胞在不同生长阶段脂质的动态变化。我们进一步讨论了粟酒裂殖酵母Deltaplh1Deltadga1细胞程序性细胞死亡过程中磷脂、鞘脂和中性脂质的动态变化。
