title: Neutral glycosphingolipids in human blood: a precise mass spectrometry analysis with special reference to lipoprotein-associated Shiga toxin receptors.
abstract: Shiga toxin (Stx)-producing Escherichia coli are the leading cause of hemorrhagic colitis and life-threatening extraintestinal complications in humans. Stx1 and Stx2 are transferred by yet to be delineated mechanisms from the intestine to the circulation where they injure microvascular endothelial cells. The resulting vascular lesions cause renal failure and brain damage. Because lipoproteins are potential carriers of Stx through the circulation, we investigated human lipoprotein-associated neutral glycosphingolipids (GSLs) with emphasis on high (globotriaosylceramide) and low (globotetraosylceramide) affinity Stx-receptors. TLC overlay employing Stx1, Stx2, and anti-GSL antibodies demonstrated preferential distribution of globo-series GSLs to very low- and low-density lipoproteins compared with minor association with high-density lipoproteins. Electrospray ionization quadrupole time-of-flight mass spectrometry portrayed C24:0/C24:1 and C16:0 as the major fatty acid of the ceramide moieties of Stx-receptors carrying nonvarying d18:1 sphingosine. This structural heterogeneity was also found in precursor lactosylceramide, glucosylceramide, and galactosylceramide, the last showing an exceptionally high degree of hydroxylated C24 fatty acids. Our findings provide the basis for exploring the functional role of lipoprotein-associated Stx-receptors in human blood.
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Christian H
authors.initials: CH
authors.lastname: Schweppe
authors.forename: Petra
authors.initials: P
authors.lastname: Hoffmann
authors.forename: Jerzy-Roch
authors.initials: JR
authors.lastname: Nofer
authors.forename: Gottfried
authors.initials: G
authors.lastname: Pohlentz
authors.forename: Michael
authors.initials: M
authors.lastname: Mormann
authors.forename: Helge
authors.initials: H
authors.lastname: Karch
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Friedrich
authors.forename: Johannes
authors.initials: J
authors.lastname: Müthing
mesh_terms: D000906:Antibodies / Q000276:immunology; D001774:Blood Chemical Analysis / Q000379:methods* / Q000592:standards*; D002554:Cerebrosides / Q000097:blood; D002855:Chromatography, Thin Layer; D005732:Gangliosides / Q000097:blood; D006801:Humans; D007118:Immunoassay; D008074:Lipoproteins / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards*; D020383:Neutral Glycosphingolipids / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D012015:Reference Standards; D014281:Trihexosylceramides / Q000097:blood* / Q000737:chemistry
chemical_list: D000906:Antibodies; D002554:Cerebrosides; D005732:Gangliosides; D008074:Lipoproteins; D020383:Neutral Glycosphingolipids; D014281:Trihexosylceramides; C013870:ceramide monohexoside; C038244:ganglio-N-triaosylceramide; C018549:globotriaosylceramide
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