Mass spectrometry-based profiling of plant sphingolipids from typical and aberrant metabolism

Rebecca E. Cahoon, Ariadna Gonzalez Solis, Jennifer E. Markham, Edgar B. Cahoon

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Mass spectrometry has increasingly been used as a tool to complement studies of sphingolipid metabolism and biological functions in plants and other eukaryotes. Mass spectrometry is now essential for comprehensive sphingolipid analytical profiling because of the huge diversity of sphingolipid classes and molecular species in eukaryotes, particularly in plants. This structural diversity arises from large differences in polar head group glycosylation as well as carbon-chain lengths of fatty acids and desaturation and hydroxylation patterns of fatty acids and long-chain bases that together comprise the ceramide hydrophobic backbone of glycosphingolipids. The standard methods for liquid chromatography–mass spectrometry (LC-MS)-based analyses of Arabidopsis thaliana leaf sphingolipids profile >200 molecular species of four sphingolipid classes and free long-chain bases and their phosphorylated forms. While these methods have proven valuable for A. thaliana based sphingolipid research, we have recently adapted them for use with ultraperformance liquid chromatography separations of molecular species and to profile aberrant sphingolipid forms in pollen, transgenic lines, and mutants. This chapter provides updates to standard methods for LC-MS profiling of A. thaliana sphingolipids to expand the utility of mass spectrometry for plant sphingolipid research.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages157-177
Number of pages21
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2295
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Ceramide
  • Deoxysphinganine
  • GIPC
  • Glucosylceramide
  • Glycosylinositolphosphoceramide
  • Long-chain base
  • Serine palmitoyltransferase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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