Analysis of yeast lipid droplet proteome and lipidome

Methods Cell Biol. 2013:116:15-37. doi: 10.1016/B978-0-12-408051-5.00002-4.

Abstract

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.

Keywords: Fatty acids; Gas liquid chromatography; Lipid droplets/particles; Mass spectrometry; Nonpolar lipids; Phospholipids; Thin layer chromatography; Yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Thin Layer / methods
  • Gas Chromatography-Mass Spectrometry / methods
  • Inclusion Bodies / chemistry
  • Inclusion Bodies / metabolism*
  • Lipid Metabolism*
  • Lipids / chemistry
  • Lipids / isolation & purification*
  • Mass Spectrometry / methods
  • Pichia
  • Proteomics / methods*
  • Saccharomyces cerevisiae
  • Yarrowia

Substances

  • Lipids