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Glycobiology Advance Access published online on April 13, 2006
Glycobiology, doi:10.1093/glycob/cwj114
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© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received January 31, 2006
Revised April 6, 2006
Accepted April 7, 2006

Article

Isolation and identification of nine sulfated glycosphingolipids containing two unique sulfated gangliosides from the African green monkey kidney cells, Verots S3, and their possible metabolic pathways

Yukio Niimura 1 * and Ineo Ishizuka 2

1 Research Center of Biomedical Analysis and Radioisotope, Teikyo University School of Medicine, 2-11-1 Kaga Itabashi-ku, Tokyo 173-8605, Japan
2 Department of Biochemistry, Teikyo University School of Medicine, 2-11-1 Kaga Itabashi-ku, Tokyo 173-8605, Japan

* To whom correspondence should be addressed.
Yukio Niimura, E-mail: yniimura{at}med.teikyo-u.ac.jp


  Abstract

Verots S3 cells derived from the African green monkey kidney were revealed to contain nine types of sulfoglycolipidsby incorporati ng [35S]sulfate. These sulfated glycolipids were separated by DEAE-Sephadex column chromatography and preparative thin-layer chromatography. The major sulfoglycolipids were characterized by using thin-layer chromatography, gas-liquid chromatography, mass spectrometry, solvolysis, thin-layer chromatography immunostaining, and nuclear magnetic resonance spectra as follows:V1 , SM4s (GalCer I3-sulfate);V2 ,SM3 (LacCer II3-sulfate);V3 ,SM2a (Gg3Cer II3-sulfate); V4, globopentaosyl ceramide sulfate (Gb5Cer V3-sulfate); V5, (Gg4Cer II3-sulfate, IV3-NeuAc); V6, SB1a (Gg4Cer II3, IV3-bis-sulfate); and V8, (Gg4Cer II3-NeuAc, IV3-sulfate). Both V5 and V8 were sulfated gangliosides comprising both N-acetyl neuraminic acid and sulfate, and this was the first report on V8. A minor component V7 was identified as SM1a (Gg4Cer II3-sulfate) based on its behavior in thin-layer chromatography, gas-liquid chromatography, and liquid secondary ion mass spectroscopy. It was postulated that this substance was a precursor of V6 (SB1a) and V5 (Gg4Cer II3-sulfate, IV3-NeuAc) and to date, its presence has not been demonstrated in nature. Anotherm inor component V9 was identifieda s glucosyl ceramide sulfate based on itsm igration in thin-layer chromatography and gas-liquid chromatography. This renal cell line was shown to be an excellent model for studying the metabolism and function of sulfoglycolipids.

Keywords: African green monkey kidney cell/biosynthetic pathways of sulfoglycolipids/sulfated ganglioside/sulfoglycolipids.
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