title: Verification and refinement of cellular glycosphingolipid profiles using HPLC.
abstract: Glycosphingolipids (GSLs) are hybrid molecules consisting of the sphingolipid ceramide linked to a mono- or oligo-saccharide. In comparison to other membrane lipids, the family of GSLs stands out because of the extensive variation in the carbohydrate headgroup. GSLs are cell surface binding partners, in cis with growth factor receptors, and in trans with bacterial toxins and viruses, and are among the host-derived membrane components of viral particles, including those of HIV. In spite of their biological relevance, GSL profiles of commonly used cell lines have been analyzed to different degrees. Here, we directly compare the GSL complements from CHO-K1, COS-7, HeLa, HEK-293, HEPG2, Jurkat, and SH-SY5Y cells using an HPLC-based method requiring modest amounts of material. Compared to previous studies, the HPLC-based analyses provided more detailed information on the complexity of the cellular GSL complement, qualitatively as well as quantitatively. In particular for cells expressing multiple GSLs, we found higher numbers of GSL species, and different levels of abundance. Our study thus extends our knowledge of biologically relevant lipids in widely used cell lines.
authors.affiliation: a Atlantic Research Centre, Departments of Pediatrics and Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
authors.forename: Douglas B
authors.initials: DB
authors.lastname: Vieira
authors.affiliation: a Atlantic Research Centre, Departments of Pediatrics and Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
authors.forename: Karen
authors.initials: K
authors.lastname: Thur
authors.affiliation: a Atlantic Research Centre, Departments of Pediatrics and Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
authors.forename: Saki
authors.initials: S
authors.lastname: Sultana
authors.affiliation: b Department of Pharmacology, University of Oxford, Oxford, OX1 3QT, UK.
authors.forename: David
authors.initials: D
authors.lastname: Priestman
authors.affiliation: a Atlantic Research Centre, Departments of Pediatrics and Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
authors.forename: Aarnoud C
authors.initials: AC
authors.lastname: van der Spoel
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000378:metabolism; D006028:Glycosphingolipids / Q000737:chemistry / Q000378:metabolism*; D047709:Hirudo medicinalis / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D055432:Metabolomics / Q000379:methods; D051379:Mice; D055616:Microtechnology / Q000379:methods; D015394:Molecular Structure; D051381:Rats; D013050:Spectrometry, Fluorescence; D062367:ortho-Aminobenzoates / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D006028:Glycosphingolipids; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D006026:Glycoside Hydrolases; C051111:ceramide glycanase
keywords: HPLC; ceramide glycanase; glycosphingolipids; lipidomics; quantitative analysis