Specificities of N-acetylglucosamine-6-O-sulfotransferases in relation to L-selectin ligand synthesis and tumor-associated enzyme expression

• Published in: The Journal of biological chemistry Vol. 277; no. 6; pp. 3979 - 3984
• Main Authors: Uchimura, Kenji, El-Fasakhany, Fathy M, Hori, Mayuko, Hemmerich, Stefan, Blink, Sarah E, Kansas, Geoffrey S, Kanamori, Akiko, Kumamoto, Kensuke, Kannagi, Reiji, Muramatsu, Takashi
• Format: Journal Article
• Language: English
• Published: United States 08-02-2002
• Subjects: Base Sequence; Carbohydrate Sulfotransferases; Colonic Neoplasms - enzymology; Colonic Neoplasms - pathology; DNA Primers; Humans; Isoenzymes - metabolism; L-Selectin - metabolism; Oligosaccharides - chemistry; Oligosaccharides - metabolism; Substrate Specificity; Sulfotransferases - genetics; Sulfotransferases - metabolism; Transfection
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M106587200

N-Acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) catalyzes the transfer of sulfate from adenosine 3'-phosphate,5'-phosphosulfate to the C-6 position of the non-reducing GlcNAc. Three human GlcNAc6STs, namely GlcNAc6ST-1, GlcNAc6ST-2 (HEC-GlcNAc6ST), and GlcNAc6ST-3 (I-GlcNAc6ST), were produced as fusion proteins to protein A, and their substrate specificities as well as their enzymological properties were determined. Both GlcNAc6ST-1 and GlcNAc6ST-2 efficiently utilized the following oligosaccharide structures as acceptors: GlcNAcbeta1-6[Galbeta1-3]GalNAc-pNP (core 2), GlcNAcbeta1-6ManOMe, and GlcNAcbeta1-2Man. The ratios of activities to these substrates were not significantly different between the two enzymes. However, GlcNAc6ST-2 but not GlcNAc6ST-1 acted on core 3 of GlcNAcbeta1-3GalNAc-pNP. GlcNAc6ST-3 used only the core 2 structure among the above mentioned oligosaccharide structures. The ability of GlcNAc6ST-1 to sulfate core 2 structure as efficiently as GlcNAc6ST-2 is consistent with the view that GlcNAc6ST-1 is also involved in the synthesis of l-selectin ligand. Indeed, cells doubly transfected with GlcNAc6ST-1 and fucosyltransferase VII cDNAs supported the rolling of L-selectin-expressing cells. The activity of GlcNAc6ST-2 on core 3 and its expression in mucinous adenocarcinoma suggested that this enzyme corresponds to the sulfotransferase, which is specifically expressed in mucinous adenocarcinoma (Seko, A., Sumiya, J., Yonezawa, S., Nagata, K., and Yamashita, K. (2000) Glycobiology 10, 919-929).