High-content screening of yeast mutant libraries by shotgun lipidomics

被引：19

作者：

Tarasov, Kirill ^{[1
,2
]}

Stefanko, Adam ^{[3
,4
]}

Casanovas, Albert ^{[3
]}

Surma, Michal A. ^{[4
]}

Berzina, Zane ^{[3
]}

Hannibal-Bach, Hans Kristian ^{[3
]}

Ekroos, Kim ^{[1
]}

Ejsing, Christer S. ^{[3
]}

机构：

[1] Zora Biosci Oy, FI-02150 Espoo, Finland

[2] Univ Montreal, Dept Biochem & Mol Med, Montreal, PQ H3T 1J4, Canada

[3] Univ Southern Denmark, Dept Biochem & Mol Biol, DK-5230 Odense, Denmark

[4] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany

来源：

MOLECULAR BIOSYSTEMS | 2014年 / 10卷 / 06期

关键词：

SACCHAROMYCES-CEREVISIAE; MEMBRANE-PROTEINS; SPHINGOLIPID METABOLISM; PHOSPHOLIPID-SYNTHESIS; SECRETORY VESICLES; GLOBAL ANALYSIS; LIPIN HOMOLOG; GENE; TRANSCRIPTION; REVEALS;

D O I：

10.1039/c3mb70599d

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

To identify proteins with a functional role in lipid metabolism and homeostasis we designed a high-throughput platform for high-content lipidomic screening of yeast mutant libraries. To this end, we combined culturing and lipid extraction in 96-well format, automated direct infusion nanoelectrospray ionization, high-resolution Orbitrap mass spectrometry, and a dedicated data processing framework to support lipid phenotyping across hundreds of Saccharomyces cerevisiae mutants. Our novel approach revealed that the absence of genes with unknown function YBR141C and YJR015W, and the transcription factor KAR4 precipitated distinct lipid metabolic phenotypes. These results demonstrate that the high-throughput shotgun lipidomics platform is a valid and complementary proxy for high-content screening of yeast mutant libraries.

引用

页码：1364 / 1376

页数：13

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