Abstract
A novel sulfated glycosphingolipid containing a sulfated glucosyl residue was isolated from rat kidney and purified to homogeneity by column chromatographies with DEAE-Sephadex and silica beads. By compositional analyses, permethylation studies, one- and two-dimensional proton magnetic resonance spectroscopy, infrared spectroscopy, negative secondary ion mass spectrometry, solvolysis, and immunostaining on thin layer chromatogram, the structure of this glycolipid was proposed to be HSO3-3Glc beta 1-1Cer (where Cer is ceramide). The ceramide portion consisted of 4-D-hydroxysphinganine as the sole long chain base, and the fatty acid consisted of predominantly tetracosanoic acid, deduced from both composition analysis and negative secondary ion mass spectrometry. The yield of glucosyl sulfatide was about 5 nmol/g of tissue, being about three times as much as that of lactosylceramide sulfate.
Highlights
A novel sulfated glycosphingolipid containing a sul- contain a novel sulfatide, GlcCer sulfate, whichcould not be separated from SM4s on TLC and purified to homogeneity by column chromatogra- unless an acidic solvent system was used
The ceramide portion consisted of 4D-hydroxysphinganineas the sole long chain base, and the fatty acid consisted of predominantlytetracosanoic acid, deduced from both composition analysis and neg
HCl/water (82:8.6:9.4, v/v) at 70 "C for 18 h [25], treated with week-old Wistar rats were extracted in three steps with chlomethanol/acetic anhydride (41, v/v) (261, and incubated with BSTFA/pyridine (9:1, v/v) to obtain the N-acetyl-0-trimethylsilyl derivatives of sphingoid bases
Summary
SM2, gangliotriaosylceramide monosulfate (GgOsesCer I13-sulfate); Thin Layer Chromatography-TLC was performed on Silica Gel. Lac, D-lactose; Cer, ceramide; SM45, galactosylceramide sulfate 60 HPTLC plates with the following solvent systems: I, chloroform/. The "C at the rateof 4 "C/min.Analyses of the methyl esters of unsatu- spectra were obtained by a GX-400 spectrometer of Japan Electron rated fatty acids were achieved by using a glass column (3.2 mm X Optical Laboratory (JEOL) equipped with a JEOL PLEXUS data. Gas Chromatography-Mass Spectrometry-GC-MS was performed times; temperature, 30 "C. by a Shimadzu model 6020 Auto GC-MS apparatus equipped with a Two-dimensional multiple relayed COSY spectra were obtained by SCAP 1123data system using a glass column (3.2 mm X 1m) of 3% using the pulse sequences of Bax of Drobny [33]. The acetate of partially methylated alditol was analyzed by GLC and GC-MS as described above
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