Mono-sulfated Globopentaosylceramide from Human Kidney

Abstract

A novel sulfated glycosphingolipid that belongs to the "globo-series" was isolated from human kidney. This lipid was purified from a pooled kidney preparation by chloroform/methanol extraction, mild alkaline treatment, DEAE-Sephadex and silicic acid column chromatographies, and preparative thin layer chromatography. The structure and the properties were studied by infrared spectroscopy, two-dimensional proton magnetic resonance spectroscopy, negative secondary ion mass spectrometry, solvolysis, compositional and methylation analyses, monoclonal antibodies, and sulfatide-binding proteins. From the results of the above analyses, the structure of this glycolipid was proposed to be HSO3-3Gal beta 1-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1-1ceramide. The proton resonance at 3.93 ppm of the H-3 of the sulfated nonreducing terminal galactose of this lipid was downfield-shifted (delta 0.48 ppm), as compared with H-3 of the internal beta-galactose because of the electronegativity of the sulfate ester. This sulfated lipid reacted with a monoclonal anti-SSEA-3 (MC-631) (Kannagi, R., Cochran, N. A., Ishigami, F., Hakomori, S., Andrews, P. W., Knowles, B. B., and Solter, D. (1983) EMBO J. 2, 2355-2361), whose epitope is R-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-R', on thin layer chromatograms and solid-phase radioimmunoassay. This lipid also bound to the 125I-labeled sulfatide-binding protein, thrombospondin. The yield of this sulfated glycolipid was 0.19 nmol/g of tissue, which was about 0.09 and 0.5 mol % of galactosyl and lactosyl sulfatides in human kidney.