Molecular cloning, sequence, and expression of a human GDP-L-fucose:beta-D-galactoside 2-alpha-L-fucosyltransferase cDNA that can form the H blood group antigen

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摘要：

We have previously used a gene-transfer scheme to isolate a human genomic DNA fragment that determines expression of a GDP-L-fucose:β-D-galactoside 2-α-L-fucosyltransferase [α(1,2)FT; EC 2.4.1.69]. Although this fragment determined expression of an α(1, 2)FT whose kinetic properties mirror those of the human H blood group α(1,2)FT, their precise nature remained undefined. We describe here the molecular cloning, sequence, and expression of a human cDNA corresponding to these human genomic sequences. When expressed in COS-1 cells, this cDNA directs expression of cell surface H structures and a cognate α(1,2)FT activity with properties analogous to the human H blood group α(1,2)FT. The cDNA sequence predicts a 365-amino acid polypeptide characteristic of a type II transmembrane glycoprotein with a domain structure analogous to that of other glycosyltransferases but without significant primary sequence similarity to these or other known proteins. To directly demonstrate that the cDNA encodes an α(1,2)FT, the COOH-terminal domain predicted to be Golgi-resident was expressed in COS-1 cells as a catalytically active, secreted, and soluble protein A fusion peptide. Southern blot analysis showed that this cDNA identifies DNA sequences syntenic to the human H locus on chromosome 19. These results strongly suggest that this cloned α(1,2)FT cDNA represents the product of the human H blood group locus.

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关键词：

Biochemistry Oligosaccharide Biosynthesis Glycosyltransferase Surface Antigen Chromosome 19

DOI：

10.1073/pnas.87.17.6674

被引量：

700

年份：

1990