Molecular characterization of a fucosyltransferase encoded by Schistosoma mansoni.
The glycans of schistosomes include many complex carbohydrates that contain fucose. Although the biological functions of these complex carbohydrates are not yet clearly understood, some of these structures are thought to play essential roles in the life cycle of the parasite. Here we present the molecular cloning and characterization of a fucosyltransferase of Schistosoma mansoni with a DNA sequence similarity of 84.6 and 63.7% to mouse and human fucosyltransferase type VII. Southern blot analysis of genomic DNA indicated that this S. mansoni fucosyltransferase is the product of a single gene. The schistosome cDNA sequence that we obtained contains an open reading frame encoding a protein of 351 amino acids with a predicted molecular size of 40.5 kDa. From the amino acid sequence, we predicted two potential N-linked and one O-linked glycosylation site. Western blot studies of extracts from stably transfected CHO cells showed a band corresponding to the schistosome fucosyltransferase at 50 kDa, suggesting that the enzyme is indeed glycosylated. We further demonstrated the expression and enzymatic activity of the fucosyltransferase in the transfected cells by immunofluorescence studies and flow microfluorimetric analysis, which indicated that the enzyme is capable of synthesizing the SLeX blood group determinant but not the LeX determinant in CHO cells. The identification of a fucosyltransferase type VII in schistosomes further underscores the importance of fucose-containing glycans in schistosome glycobiology.
Marques
E T
ET
Johns Hopkins University School of Medicine, Department of Pharmacology and Molecular Science, Baltimore, MD 21205, USA. emarques@welchlink.welch.jhu.edu
Weiss
J B
JB
Strand
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D005647:Fucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017238:Genes, Helminth; D006801:Humans; D016256:Lewis X Antigen / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000096:biosynthesis; D016366:Open Reading Frames; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics; D000080506:Sialyl Lewis X Antigen; D014162:Transfection
D018076:DNA, Complementary; D016256:Lewis X Antigen; D009844:Oligosaccharides; D000080506:Sialyl Lewis X Antigen; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase