title: Database and data analysis application for structural characterization of gangliosides and sulfated glycosphingolipids by negative ion mass spectrometry.
abstract: Gangliosides and sulfated glycosphingolipids, as building and functional components of animal cell membranes, participate in cell-to-cell interactions and signaling, but also in changes of cell architecture due to different pathophysiological events. In order to enable higher throughput and to facilitate structural characterization of gangliosides/sulfo-glycosphingolipids (GSL) and their neutral GSL counterparts by negative ion mass spectrometry (MS) and tandem MS techniques, a database and data analysis application have been developed. In silico developed glycosphingolipid database considers a high diversity of ceramide compositions, several sialic acid types (N-acetylneuraminic acid, N-glycolylneuraminic acid and 2-keto-3-deoxynononic acid) as well as possible additional substitutions/modifications of glycosphingolipids, such as O-acetylation, de-N-acetylation, fucosylation, glucuronosylation, sulfation, attachment of repeating terminal hexose-N-acetylhexosamine- (Hex-HexNAc-)1-6 extension, and possible lactone forms. Data analysis application, named GSL-finder, enables correlation of negative ion MS and/or low-energy tandem MS spectra with the database structures. The GSL-database construction and the GSL-finder application searching rules are explained. Validation conducted on GD1a fraction as well as on complex mixtures of native gangliosides isolated from different mammalian brain tissues (human fetal and adult brain, and calf brain tissue) demonstrated agreement with previous studies. Plain, fast, and automated routine for structural characterization of gangliosides/sulfated glycosphingolipids and their neutral GSL counterparts described here could facilitate and improve mass spectrometric analysis of complex glycosphingolipid mixtures originating from variety of normal and pathological biomaterial, where it is known that distinctive changes in glycosphingolipid composition occur.
authors.affiliation: Department of Physical Chemistry, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia. Electronic address: marko@irb.hr.
authors.forename: Marko
authors.initials: M
authors.lastname: Rožman
authors.affiliation: Department for Chemistry and Biochemistry, School of Medicine, University of Zagreb, Šalata 3, 10000 Zagreb, Croatia. Electronic address: dragana.fabris@mef.hr.
authors.forename: Dragana
authors.initials: D
authors.lastname: Fabris
authors.affiliation: Department of Physical Chemistry, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia. Electronic address: tomislav.mrla@irb.hr.
authors.forename: Tomislav
authors.initials: T
authors.lastname: Mrla
authors.affiliation: Department for Chemistry and Biochemistry, School of Medicine, University of Zagreb, Šalata 3, 10000 Zagreb, Croatia. Electronic address: zvukelic@mef.hr.
authors.forename: Željka
authors.initials: Ž
authors.lastname: Vukelić
mesh_terms: D000818:Animals; D002417:Cattle; D002518:Ceramides / Q000737:chemistry / Q000378:metabolism; D003198:Computer Simulation; D062126:Databases, Chemical; D005732:Gangliosides / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D006028:Glycosphingolipids / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D006801:Humans; D012794:Sialic Acids / Q000737:chemistry / Q000378:metabolism; D013431:Sulfates / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D002518:Ceramides; D005732:Gangliosides; D006028:Glycosphingolipids; D012794:Sialic Acids; D013431:Sulfates
keywords: Database; Glycosphingolipid analysis; MS ion search; MSMS ion search