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@ARTICLE{vonGerichten:124296,
author = {J. von Gerichten$^*$ and K. Schlosser and D. Lamprecht$^*$
and I. Morace$^*$ and M. Eckhardt and D. Wachten and R.
Jennemann$^*$ and H.-J. Gröne$^*$ and M. Mack and R.
Sandhoff$^*$},
title = {{D}iastereomer-specific quantification of bioactive
hexosylceramides from bacteria and mammals.},
journal = {Journal of lipid research},
volume = {58},
number = {6},
issn = {1539-7262},
address = {Bethesda, Md.},
publisher = {ASBMB},
reportid = {DKFZ-2017-01192},
pages = {1247 - 1258},
year = {2017},
abstract = {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.},
cin = {G131 / G130},
ddc = {540},
cid = {I:(DE-He78)G131-20160331 / I:(DE-He78)G130-20160331},
pnm = {317 - Translational cancer research (POF3-317)},
pid = {G:(DE-HGF)POF3-317},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:28373486},
pmc = {pmc:PMC5454501},
doi = {10.1194/jlr.D076190},
url = {https://inrepo02.dkfz.de/record/124296},
}