2136–2141,PNAS,February 17, 2009,vol. 106,no. 7

【abstract】

　　Although the transcriptome, proteome, and interactome of several eukaryotic model organisms have been described in detail, lipidomes remain relatively uncharacterized. Using Saccharomyces cerevisiae as an example,wedemonstrate that automated shotgun lipidomics analysis enabled lipidome-wide absolute quantification of individual molecular lipid species by streamlined processing of a single sample of only 2 million yeast cells. By comparative lipidomics, we achieved the absolute quantification of 250 molecular lipid species covering 21 major lipid classes. This analysis provided 95% coverage of the yeast lipidome achieved with 125-fold improvement in sensitivity compared with previous approaches.

       Comparative lipidomics demonstrated that growth temperature and defects in lipid biosynthesis induce ripple effects throughout the molecular composition of the yeast lipidome. This work serves as a resource for molecular characterization of eukaryotic lipidomes, and establishes shotgun lipidomics as a powerful platform for complementing biochemical studies and other systems-level approaches.

Mass Spectrometry. Lipid extracts were analyzed in negative and positive ion mode on a QSTAR Pulsar-i instrument (MDS Analytical Technologies) and a LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific) both equipped with the robotic nanoflow ion source TriVersa NanoMate (Advion Biosciences).