Glycobiology, 2002, Vol. 12, No. 3 153-162
© 2002 Oxford University Press
Biosynthesis of the carbohydrate antigenic determinants, Globo H, blood group H, and Lewis b: a role for prostate cancer cell 
1,2-L-fucosyltransferase
Molecular and Cellular Biophysics, Roswell Park Cancer Institute, Buffalo, NY 14263, USA
Prostate carcinoma LNCaP cells were unique among several human cancer cell lines which include two other prostate cancer cell lines, PC-3 and DU-145, in expressing 
1,2-L-fucosyltransferase (FT) as an exclusive FT activity. Affinity gel-GDP and Sephacryl S100 HR columns were used for a partial purification of this enzyme from 3.9 x 109 LNCaP cells (~200-fold; 40% yield). The Km value (2.7 mM) for the LacNAc type 2 acceptor was quite similar to the one reported for the cloned blood group H gene-specified 1,2-FT [Chandrasekaran et al. (1996) Biochemistry 35, 8914–8924]. N-Ethylmaleimide was a potent inhibitor (Ki 12.5 µM). The enzyme showed four-fold acceptor preference for the LacNAc type 2 unit in comparison to the T-hapten in mucin core 2 structure. Its main features were similar to those of the cloned enzyme: (1) C-6 sulfation of terminal Gal in the LacNAc unit increased the acceptor efficiency, whereas C-6 sialylation abolished acceptor ability; (2) C-6 sulfation of GlcNAc in LacNAc type 2 decreased by 80% the acceptor ability, whereas LacNAc type 1 was unaffected; (3) Lewis x did not serve as an acceptor; (4) the C-4 hydroxyl rather than the C-6 hydroxyl group of the GlcNAc moiety in LacNAc type1 was essential for activity; and (5) the acrylamide copolymer of Galß1,3GlcNAcß-O-Al was the best acceptor among the acrylamide copolymers. Additionally, highly significant biological features of 1,2FT were identified in the present study. The synthesis of Globo H and Lewis b determinants became evident from the fact that Galß1,3GalNAcß1,3Gal-O-Me and Galß1,3(Fuc1,4)Glc-NAcß1,3Galß-O-Me served as high-affinity acceptors for this enzyme. Further, D-Fucß1,3Gal-NAcß1,3Gal-O-Me was a very efficient acceptor, indicating that the C-6 hydroxyl group of the terminal Gal moiety in Globo H is not essential for the enzyme activity. Thus, the present study was able to demonstrate three different catalytic roles of LNCaP 1,2-FT, namely, the expressions of blood group H, Lewis b from Lewis a, and Globo H.
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