Richard Vincent R, Bourque Simon D, Titorenko Vladimir I
Department of Biology, Concordia University, 7141 Sherbrooke Street, West, SP Building, Montreal, QC, Canada, H4B 1R6.
Methods Mol Biol. 2014;1205:359-73. doi: 10.1007/978-1-4939-1363-3_21.
Metabolomic and lipidomic analyses of yeast cells provide comprehensive empirical datasets for unveiling mechanisms underlying complex biological processes. In this chapter, we describe detailed protocols for using such analyses to study the age-related dynamics of changes in intracellular and extracellular levels of various metabolites and membrane lipids in chronologically aging yeast. The protocols for the following high-throughput analyses are described: (1) microanalytic biochemical assays for monitoring intracellular concentrations of trehalose and glycogen; (2) gas chromatographic quantitative assessment of extracellular concentrations of ethanol and acetic acid; and (3) mass spectrometric identification and quantitation of the entire complement of cellular lipids. These protocols are applicable to the exploration of the metabolic patterns associated not only with aging but also with many other vital processes in yeast. The described here methodology complements the powerful genetic approaches available for mechanistic studies of fundamental aspects of yeast biology.
酵母细胞的代谢组学和脂质组学分析为揭示复杂生物过程背后的机制提供了全面的实证数据集。在本章中，我们描述了详细的实验方案，用于利用此类分析研究时序老化酵母中各种代谢物和膜脂的细胞内和细胞外水平随年龄变化的动态。描述了以下高通量分析的实验方案：（1）用于监测海藻糖和糖原细胞内浓度的微量分析生化测定；（2）气相色谱法定量评估乙醇和乙酸的细胞外浓度；（3）细胞脂质全组分的质谱鉴定和定量。这些实验方案不仅适用于探索与衰老相关的代谢模式，也适用于探索酵母中许多其他重要过程的代谢模式。此处描述的方法补充了可用于酵母生物学基本方面机制研究的强大遗传方法。
