Beta1,3-galactosyltransferase beta3Gal-T5 acts on the GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-R sugar chains of carcinoembryonic antigen and other N-linked glycoproteins, and is down-regulated in colon adenocarcinomas

doi:10.1074/jbc.M006662200

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Beta1,3-galactosyltransferase beta3Gal-T5 acts on the GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-R sugar chains of carcinoembryonic antigen and other N-linked glycoproteins, and is down-regulated in colon adenocarcinomas

Roberta Salvini, Anna Bardoni, Maurizia Valli, and Marco Trinchera

Department of Biochemistry, University of Pavia, Pavia 27100

Corresponding Author: dbioc{at}unipv.it

We attempted to determine if $beta$ 1,3galactosyltransferase $beta$ 3Gal-T5 is involved in the biosynthesis of a specific subset of type 1 chain carbohydrates and expressed in a cancer associated manner. We transfected CHO cells expressing Fuc-TIII with $beta$ 3Gal-T cDNAs and studied the relevant glycoconjugates formed. $beta$ 3Gal-T5 directs synthesis of Lewis type 1 antigens in CHO cells more efficiently than $beta$ 3Gal-T1, while $beta$ 3Gal-T2, -T3, and -T4 are almost unable. In the clone expressing Fuc-TIII and $beta$ 3Gal-T5 (CHO-FT-T5), sialyl-Lewis a synthesis is strongly inhibited by swainsonine but not by benzyl- $alpha$ -GalNAc, and sialyl-Lewis x is absent, while it is detected in the clone expressing Fuc-TIII and $beta$ 3Gal-T1 (CHO-FT-T1)or Fuc-TIII and $beta$ 3Gal-T2 (CHO-FT-T2). Endo- $beta$ -galactosidase treatment of N-glycans prepared from clone CHO-FT-T5 releases (±NeuAc $alpha$ 2-3)Gal $beta$ 1-3[Fuc $alpha$ 1-4]GlcNAc $beta$ 1-3Gal, but not GlcNAc $beta$ 1-3Gal or type 2 chain oligosaccharides, which are found in CHO-FT-T1 cells. This result indicates that $beta$ 3Gal-T5 expression prevents poly-N-acetyllactosamine and sialyl-Lewis x synthesis on N-glycans. Kinetic studies confirm that $beta$ 3Gal-T5 prefers acceptors having the GlcNAc $beta$ 1-3Gal end, including lactotriosylceramide, and competitive reverse transcriptase mediated-PCR shows that $beta$ 3Gal-T5 transcript is expressed in normal colon mucosa but not or poorly in adenocarcinomas. Moreover, recombinant carcinoembryonic antigen purified from a CHO clone expressing Fuc-TIII and $beta$ 3Gal-T5 reacts with anti-sialyl-Lewis a and carries type 1 chains on oligosaccharides released by endo-b-galactosidase. We conclude that $beta$ 3Gal-T5 down-regulation plays a relevant role in determining the cancer-associated glycosylation pattern of N-glycans.

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				J. Holgersson and J. Lofling Glycosyltransferases involved in type 1 chain and Lewis antigen biosynthesis exhibit glycan and core chain specificity Glycobiology, July 1, 2006; 16(7): 584 - 593. [Abstract] [Full Text] [PDF]

				K. Miyazaki, K. Ohmori, M. Izawa, T. Koike, K. Kumamoto, K. Furukawa, T. Ando, M. Kiso, T. Yamaji, Y. Hashimoto, et al. Loss of Disialyl Lewisa, the Ligand for Lymphocyte Inhibitory Receptor Sialic Acid-Binding Immunoglobulin-Like Lectin-7 (Siglec-7) Associated with Increased Sialyl Lewisa Expression on Human Colon Cancers Cancer Res., July 1, 2004; 64(13): 4498 - 4505. [Abstract] [Full Text] [PDF]

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