Defining the Ceramide Composition of Bovine and Human Milk Gangliosides by Direct Infusion ESI-CID Tandem Mass Spectrometry of Native and Permethylated Molecular Species

O Tara Liyanage, Chaoshuang Xia, Silvia Ringler, Bernd Stahl, Catherine E Costello*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Gangliosides are glycosphingolipids composed of an oligosaccharide that contains one or more sialic acid residues and is linked to a ceramide, a lipid composed of a long chain base (LCB) that bears an amide-linked fatty acyl group (FA). The ceramide portions of gangliosides are embedded in cell membranes; the exposed glycans interact with the extracellular environment. Gangliosides play a myriad of roles in activities such as cell-cell communication, formation of lipid rafts, cellular adhesion, calcium homeostasis, host-pathogen interaction, and viral invasion. Although the epitopes responsible for the interactions of gangliosides are located in the glycan, the epitope presentation is strongly influenced by the orientation of the attached ceramide within the lipid membrane, a feature that depends on the details of its structure, that is, the specific LCB and FA. Since the identities of both the glycan and the ceramide affect the activity of gangliosides, it is important to characterize the individual intact molecular forms. We report here a mass spectrometry-based method that combines the information gained from low-energy collision-induced dissociation (CID) measurements for the determination of the glycan with tandem mass spectra obtained at stepped higher-energy CID for the detailed characterization of the LCB and FA components of intact gangliosides. We provide results from applications of this method to the analysis of gangliosides present in bovine and human milk in order to demonstrate the assignment of LCB and FA for intact gangliosides and differential detection of isomeric ceramide structures.

Original languageEnglish
Pages (from-to)16465-16473
Number of pages9
JournalAnalytical Chemistry
Volume95
Issue number45
DOIs
Publication statusPublished - 14 Nov 2023

Keywords

  • Animals
  • Cattle
  • Ceramides/analysis
  • Gangliosides/analysis
  • Humans
  • Milk, Human/chemistry
  • Polysaccharides
  • Tandem Mass Spectrometry

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