title: Distribution of C16:0, C18:0, C24:1, and C24:0 sulfatides in central nervous system lipid rafts by quantitative ultra-high-pressure liquid chromatography tandem mass spectrometry.
abstract: Sulfated galactosylceramides (sulfatides) are glycosphingolipids associated with cholesterol- and sphingolipid-enriched membrane microdomains (lipid rafts) and are highly expressed in brain tissue. Although it is known that sulfatide species show heterogeneity in their fatty acid acyl group composition throughout brain development, their lipid raft distribution and biological relevance is poorly understood. We validated a fast and sensitive ultra-high-pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method to measure developmentally regulated sulfatide species (C16:0, C18:0, C24:1, and C24:0) in central nervous system (CNS) lipid rafts isolated without using detergent. Our UHPLC-MS/MS assay showed good accuracy and precision with a linear range of 5 to 1,000 nM for C18:0 and C24:1 sulfatides and 10 to 1,000 nM for C16:0 and C24:0 sulfatides. We applied this quantitative analysis to detergent-free lipid rafts isolated from wild-type mice and arylsulfatase A-deficient (ASA knockout) mice that accumulate sulfatides. All four sulfatide species were more abundant in raft membranes than in non-raft membranes, with a significant increase in lipid rafts isolated from ASA knockout mice. This is the first description of an analytical method to study these sulfatide species in raft and non-raft membranes and has the potential to be applied to preparations from other tissues.
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors.forename: Ana Lis
authors.initials: AL
authors.lastname: Moyano
authors.affiliation: Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors.forename: Guannan
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors.forename: Aurora
authors.initials: A
authors.lastname: Lopez-Rosas
authors.affiliation: Department of Clinical Chemistry, The Sahlgrenska Academy, University of Gothenburg, SE-413 45 Gothenburg, Sweden.
authors.forename: Jan-Eric
authors.initials: JE
authors.lastname: Månsson
authors.affiliation: Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors.forename: Richard B
authors.initials: RB
authors.lastname: van Breemen
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA. Electronic address: mgivogri@uic.edu.
authors.forename: Maria Irene
authors.initials: MI
authors.lastname: Givogri
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002553:Cerebroside-Sulfatase / Q000502:physiology*; D002784:Cholesterol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D021962:Membrane Microdomains / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D013433:Sulfoglycosphingolipids / Q000032:analysis*; D053719:Tandem Mass Spectrometry
chemical_list: D013433:Sulfoglycosphingolipids; D002784:Cholesterol; D002553:Cerebroside-Sulfatase
keywords: Membrane microdomains; Metachromatic leukodystrophy; Sulfated galactosylceramides; Tandem mass spectrometry