Schmidt Claudia, Ploier Birgit, Koch Barbara, Daum Günther
Institute of Biochemistry, Graz University of Technology, Graz, Austria.
Methods Cell Biol. 2013;116:15-37. doi: 10.1016/B978-0-12-408051-5.00002-4.
Lipid droplets (LD) are in the spotlight of lipid research because of the link of lipid storage to health and disease and the just incipient understanding of their involvement in cellular processes apart from nonpolar lipid metabolism. Yeast is an excellent model organism to study the lipidome and proteome of LD under different environmental conditions and to address new aspects of LD biology and chemistry. In this chapter, we describe a versatile protocol for the isolation of LD at high purity and address specific demands for handling different yeast species. Moreover, we discuss the analysis of the LD proteome and lipidome based on standard methods such as thin layer chromatography (TLC), gas liquid chromatography (GLC), mass spectrometry (MS) as well as GLC/MS. Finally, we point out similarities and disparities of LD proteome and lipidome from the three different yeasts Saccharomyces cerevisiae, Yarrowia lipolytica, and Pichia pastoris.
脂滴（LD）处于脂质研究的焦点，这是因为脂质储存与健康和疾病相关，而且人们刚刚开始了解它们除了参与非极性脂质代谢之外，还在细胞过程中发挥作用。酵母是一种出色的模式生物，可用于研究不同环境条件下脂滴的脂质组和蛋白质组，并探究脂滴生物学和化学的新方面。在本章中，我们描述了一种用于高纯度分离脂滴的通用方案，并阐述了处理不同酵母物种的特定要求。此外，我们还讨论了基于薄层色谱（TLC）、气相色谱（GLC）、质谱（MS）以及GLC/MS等标准方法对脂滴蛋白质组和脂质组的分析。最后，我们指出了酿酒酵母、解脂耶氏酵母和巴斯德毕赤酵母这三种不同酵母的脂滴蛋白质组和脂质组的异同。
