%0 Journal Article
%A von Gerichten, Johanna
%A Schlosser, Kerstin
%A Lamprecht, Dominic
%A Morace, Ivan
%A Eckhardt, Matthias
%A Wachten, Dagmar
%A Jennemann, Richard
%A Gröne, Hermann-Josef
%A Mack, Matthias
%A Sandhoff, Roger
%T Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals.
%J Journal of lipid research
%V 58
%N 6
%@ 1539-7262
%C Bethesda, Md.
%I ASBMB
%M DKFZ-2017-01192
%P 1247 - 1258
%D 2017
%X Mammals synthesize, cell-type specifically, the diastereomeric hexosylceramides, β-galactosylceramide (GalCer) and β-glucosylceramide (GlcCer), which are involved in several diseases, such as sphingolipidosis, diabetes, chronic kidney diseases, or cancer. In contrast, Bacteroides fragilis, a member of the human gut microbiome, and the marine sponge, Agelas mauritianus, produce α-GalCer, one of the most potent stimulators for invariant natural killer T cells. To dissect the contribution of these individual stereoisomers to pathologies, we established a novel hydrophilic interaction chromatography-based LC-MS(2) method and separated (R > 1.5) corresponding diastereomers from each other, independent of their lipid anchors. Testing various bacterial and mammalian samples, we could separate, identify (including the lipid anchor composition), and quantify endogenous β-GlcCer, β-GalCer, and α-GalCer isomers without additional derivatization steps. Thereby, we show a selective decrease of β-GlcCers versus β-GalCers in cell-specific models of GlcCer synthase-deficiency and an increase of specific β-GlcCers due to loss of β-glucoceramidase 2 activity. Vice versa, β-GalCer increased specifically when cerebroside sulfotransferase (Gal3st1) was deleted. We further confirm β-GalCer as substrate of globotriaosylceramide synthase for galabiaosylceramide synthesis and identify additional members of the human gut microbiome to contain immunogenic α-GalCers. Finally, this method is shown to separate corresponding hexosylsphingosine standards, promoting its applicability in further investigations.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:28373486
%2 pmc:PMC5454501
%R 10.1194/jlr.D076190
%U https://inrepo02.dkfz.de/record/124296