A Remodeling System of the 3′-Sulfo-Lewis a and 3′-Sulfo-Lewis x Epitopes

• Published in: The Journal of biological chemistry 2001-10, Vol.276 (42), p.38588
• Main Authors: Naoki Ikeda, Hironobu Eguchi, Shoko Nishihara, Hisashi Narimatsu, Reiji Kannagi, Tatsuro Irimura, Mitsunori Ohta, Hikaru Matsuda, Naoyuki Taniguchi, Koichi Honke
• Format: Article
• Language: English
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M107390200

It has been reported that the chemically synthesized 3′-sulfo-Le a and 3′-sulfo-Le x epitopes have a high potential as a ligand for selectins. To elucidate the physiological functions of 3′-sulfated Lewis epitopes, a remodeling system was developed using a combination of a βGal-3- O -sulfotransferase GP3ST, hitherto known α1,3/1,4-fucosyltransferases (FucT-III, IV, V, VI, VII, and IX) and arylsulfatase A. The pyridylaminated (PA) lacto- N -tetraose (Galβ1–3GlcNAcβ1–3Galβ1–4Glc) was first converted to 3′-sulfolacto- N -fucopentaose II (sulfo-3Galβ1–3(Fucα1–4)GlcNAcβ1–3Galβ1–4Glc)-PA by sequential reactions with GP3ST and FucT-III. The 3′-sulfolacto- N -fucopentaose III (sulfo-3Galβ1–4(Fucα1–3)GlcNAcβ1–3Galβ1–4Glc)-PA was then synthesized from lacto- N -neotetraose (Galβ1–4GlcNAcβ1–3Galβ1–4Glc)-PA by GP3ST and FucT-III, -IV, -V, -VI, -VII, or -IX in a similar manner. The substrate specificity for the 3′-sulfated acceptor of the α1,3-fucosyltransferases was considerably different from that for the non-substituted and 3′-sialylated varieties. When the GP3ST gene was introduced into A549 and Chinese hamster ovary cells expressing FucT-III, they began to express 3′-sulfo-Le a and 3′-sulfo-Le x epitopes, respectively, suggesting that GP3ST is responsible for their biosynthesis in vivo . The expression of the 3′-sialyl-Le x epitope on Chinese hamster ovary cells was attenuated by the introduction of GP3ST gene, indicating that GP3ST and α2,3-sialyltransferase compete for the common Galβ1–4GlcNAc-R oligosaccharides. Last, arylsulfatase A, which is a lysosomal hydrolase that catalyzes the desulfation of 3- O -sulfogalactosyl residues in glycolipids, was found to hydrolyze the sulfate ester bond on the 3′-sulfo-Le x (type 2 chain) but not that on the 3′-sulfo-Le a (type 1 chain). The present remodeling system might be of potential use as a tool for the study of the physiological roles of 3′-sulfated Lewis epitopes, including interaction with selectins.