TY  - JOUR
AU  - von Gerichten, Johanna
AU  - Schlosser, Kerstin
AU  - Lamprecht, Dominic
AU  - Morace, Ivan
AU  - Eckhardt, Matthias
AU  - Wachten, Dagmar
AU  - Jennemann, Richard
AU  - Gröne, Hermann-Josef
AU  - Mack, Matthias
AU  - Sandhoff, Roger
TI  - Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals.
JO  - Journal of lipid research
VL  - 58
IS  - 6
SN  - 1539-7262
CY  - Bethesda, Md.
PB  - ASBMB
M1  - DKFZ-2017-01192
SP  - 1247 - 1258
PY  - 2017
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:28373486
C2  - pmc:PMC5454501
DO  - DOI:10.1194/jlr.D076190
UR  - https://inrepo02.dkfz.de/record/124296
ER  -