Expression of human H-type α1,2-fucosyltransferase encoding for blood group H(O) antigen in chinese hamster ovary cells. Evidence for preferential fucosylation and truncation of polylactosamine sequences

Pedro A. Prieto, Robert D. Larsen, Moonjae Cho, Hilda N. Rivera, Ali Shilatifard, John B. Lowe, Richard D. Cummings, David F. Smith*

*Corresponding author for this work

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

The human H(O) blood group is specified by the structure Fucα1-2Galβ1- R, but the factors regulating expression of this determinant on cell surface glycoconjugates are not well understood. To learn more about the regulation of H blood group expression, cDNA encoding the human H-type GDPFuc:β-D- galactoside α1,2-fucosyltransferase (α1,2FT) was stably transfected into Chinese hamster ovary (CHO) cells. The new cell line, designated CHO(α1,2)FT, expressed surface neoglycans containing the H antigen. The structures of the fucosylated neoglycans in CHO(α1,2)FT cells and the distribution of these glycans on glycoproteins were characterized. Seventeen percent of the [ 3H]Gal-labeled glycopeptides from CHO(α1,2)FT cells bound to the immobilized H blood group-specific lectin Ulex europaeus agglutinin-I (UEA-I), whereas none from parental CHO cells bound to the lectin. The glycopeptides from CHO(α1,2)FT cells binding to UEA-I contained polylactosamine [3Galβ1-4Glc-NAcβ1-] with the terminal sequence Fucα1- 2Galβ1-4GlcNAc-R. Fucosylation of the polylactosamine sequences on complex- type N-glycans in CHO(α1,2)FT cells caused a decrease in both sialylation and length of polylactosamine. Unexpectedly, only small amounts of terminal fucosylation was found in diantennary complex-type N-glycans. The O-glycans and glycolipids were not fucosylated by the H-type α1,2FT. Two major high molecular weight glycoproteins, one of which was shown to be the lysosome- associated membrane glycoprotein LAMP-1, preferentially contained the H-type structure and were bound by immobilized UEA-I. These results demonstrate that in CHO cells the expressed H-type α1,2FT does not indiscriminately fucosylate terminal galactosyl residues in complex-type N-glycans, but it favors glycans containing polylactosamine and dramatically alters their length and sialylation.

Original languageEnglish (US)
Pages (from-to)2089-2097
Number of pages9
JournalJournal of Biological Chemistry
Volume272
Issue number4
DOIs
StatePublished - 1997

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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