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Originally published In Press as doi:10.1074/jbc.M011791200 on February 27, 2001

J. Biol. Chem., Vol. 276, Issue 20, 16695-16703, May 18, 2001
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A Novel Ganglioside Isolated from Renal Cell Carcinoma*

Akihiro ItoDagger , Steven B. Levery§, Seiichi Saito, Makoto Satoh, and Sen-itiroh HakomoriDagger ||

From the Dagger  Division of Biomembrane Research, Pacific Northwest Research Institute and the Departments of Pathobiology and Microbiology, University of Washington, Seattle, Washington 98122, the § Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602, and the  Department of Urology, Tohoku University School of Medicine, Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan

In renal cell carcinoma (RCC), the level of higher gangliosides is correlated with degree of metastatic potential, and cell lines derived from metastatic deposits of RCC are characterized by high expression of disialogangliosides (Saito, S., Orikasa, S., Ohyama, C., Satoh, M., and Fukushi, Y. (1991) Int. J. Cancer 49, 329-334 and Saito, S., Orikasa, S., Satoh, M., Ohyama, C., Ito, A., and Takahashi, T. (1997) Jpn. J. Cancer Res. (Gann) 88, 652-659). We now report two disialogangliosides, G1 and G2, found in the RCC cell line TOS-1. G1 from TOS-1 cells was characterized as having a novel hybrid structure between ganglio-series (region I as in Structure TI; same as the terminal structure of ganglioside GM2), and the lacto-series type 1 (region II). The characterization was based on reactivity with various monoclonal antibodies (mAbs) with defined epitope specificity, as well as monosaccharide and fatty acid component analysis, 1H NMR spectroscopy, and electrospray ionization mass spectrometry of the intact compound. G1 showed strong reactivity with mAb RM2, raised originally against TOS-1 cells, and weak cross-reactivity with anti-GM2 mAb MK-1-8. The antigen is hereby termed GalNAc disialosyl Lc4Cer (IV4GalNAcIV3NeuAcIII6NeuAcLc4; abbreviated GalNAcDSLc4). G2 was identified by 1H NMR and mass spectrometry as having a structure similar to Structure TI but without the GalNAcbeta 1right-arrow4 substitution and showed strong reactivity with mAb FH9 reported previously to be specific for disialosyl lacto-series type 1 (disialosyl Lc4) having vicinal alpha 2right-arrow3 and alpha 2right-arrow6 sialosyl residues, an antigen associated with human colonic cancer. Clinicopathological studies indicate that expression of these disialoganglioside antigens in RCC tissue is correlated with the metastatic potential of RCC.

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  		Structure I.  

* This study was supported by NCI, National Institutes of Health (NIH) Grants CA80054 and CA82167 (to S. H.) and the NIH Resource for Complex Biomedical Carbohydrates, P41 RR005351 (to S. B. L.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed. Tel.: 206-726-1222; Fax: 206-726-1212; E-mail: hakomori@u.washington.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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