Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S]
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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2017-06-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520310178 |
| _version_ | 1818417888890978304 |
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| author | Johanna von Gerichten Kerstin Schlosser Dominic Lamprecht Ivan Morace Matthias Eckhardt Dagmar Wachten Richard Jennemann Hermann-Josef Gröne Matthias Mack Roger Sandhoff |
| author_facet | Johanna von Gerichten Kerstin Schlosser Dominic Lamprecht Ivan Morace Matthias Eckhardt Dagmar Wachten Richard Jennemann Hermann-Josef Gröne Matthias Mack Roger Sandhoff |
| author_sort | Johanna von Gerichten |
| collection | DOAJ |
| description | 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-MS2 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. |
| first_indexed | 2024-12-14T12:13:57Z |
| format | Article |
| id | doaj.art-c86c3de3fab0410f9a9efffb263caea3 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T12:13:57Z |
| publishDate | 2017-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-c86c3de3fab0410f9a9efffb263caea32022-12-21T23:01:41ZengElsevierJournal of Lipid Research0022-22752017-06-0158612471258Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S]Johanna von Gerichten0Kerstin Schlosser1Dominic Lamprecht2Ivan Morace3Matthias Eckhardt4Dagmar Wachten5Richard Jennemann6Hermann-Josef Gröne7Matthias Mack8Roger Sandhoff9Lipid Pathobiochemistry Group German Cancer Research Center, Heidelberg, Germany; Instrumental Analytics and Bioanalytics, Mannheim University of Applied Sciences, Mannheim, GermanyDepartment of Biotechnology, Institute for Technical Microbiology, Mannheim University of Applied Sciences, Mannheim, GermanyLipid Pathobiochemistry Group German Cancer Research Center, Heidelberg, Germany; Center for Applied Research in Biomedical Mass Spectrometry (ABIMAS), Mannheim University of Applied Sciences, Mannheim, GermanyDepartment of Molecular and Cellular Pathology, German Cancer Research Center, Heidelberg, GermanyInstitute of Biochemistry and Molecular Biology and Center for Rare Diseases University of Bonn, Bonn, GermanyMinerva Max Planck Research Group, Molecular Physiology, Center of Advanced European Studies and Research, Bonn, Germany; Institute of Innate Immunity, University Hospital, University of Bonn, Bonn, GermanyDepartment of Molecular and Cellular Pathology, German Cancer Research Center, Heidelberg, GermanyDepartment of Molecular and Cellular Pathology, German Cancer Research Center, Heidelberg, GermanyDepartment of Biotechnology, Institute for Technical Microbiology, Mannheim University of Applied Sciences, Mannheim, GermanyTo whom correspondence should be addressed.; Lipid Pathobiochemistry Group German Cancer Research Center, Heidelberg, Germany; Center for Applied Research in Biomedical Mass Spectrometry (ABIMAS), Mannheim University of Applied Sciences, Mannheim, GermanyMammals 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-MS2 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.http://www.sciencedirect.com/science/article/pii/S0022227520310178hydrophilic interaction chromatographyelectrospray ionization-tandem mass spectrometryglucosylceramideglucocerebrosidegalactosylceramidecerebroside |
| spellingShingle | Johanna von Gerichten Kerstin Schlosser Dominic Lamprecht Ivan Morace Matthias Eckhardt Dagmar Wachten Richard Jennemann Hermann-Josef Gröne Matthias Mack Roger Sandhoff Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] Journal of Lipid Research hydrophilic interaction chromatography electrospray ionization-tandem mass spectrometry glucosylceramide glucocerebroside galactosylceramide cerebroside |
| title | Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] |
| title_full | Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] |
| title_fullStr | Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] |
| title_full_unstemmed | Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] |
| title_short | Diastereomer-specific quantification of bioactive hexosylceramides from bacteria and mammals[S] |
| title_sort | diastereomer specific quantification of bioactive hexosylceramides from bacteria and mammals s |
| topic | hydrophilic interaction chromatography electrospray ionization-tandem mass spectrometry glucosylceramide glucocerebroside galactosylceramide cerebroside |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520310178 |
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