Nucleic acids and proteins of a rat ganglioside GM1-specific alpha1-2fucosyltransferase and uses thereof

Abstract

A rat ganglioside GM 1 -specific α1→2fucosyltransferase is disclosed. Nucleotide sequences of a rat ganglioside GM 1 -specific α1→2fucosyltransferase, amino acid sequences of its encoded protein (including peptide or polypeptide), and derivatives thereof are described. Also described are fragments (and derivatives and analogs thereof) which comprise a domain of rat ganglioside GM 1 -specific α1→2fucosyltransferase with catalytic activity. Methods of production of rat ganglioside GM 1 -specific α1→2fucosyltransferase and derivatives and analogs thereof (e.g. by recombinant means) are provided. Methods of inhibiting the function of rat ganglioside GM 1 -specific α1→2fucosyltransferase (e.g. by means of antisense RNA) are provided. Methods of commercial scale use of the rat ganglioside GM 1 -specific α1→2fucosyltransferase in the production of fucosyl-saccharide compositions are described. Applications of these compositions, e.g. as additives for human nutritive compositions or immunotherapeutics for cancer, are disclosed.

1 . A method to suppress or inhibit in a cell the function of an α1→fucosyltransferase protein, said method comprising contacting a cell with a nucleic acid comprising a nucleotide sequence that is the reverse complement of a nucleotide sequence as depicted in  FIG. 5  (SEQ ID NO:7) or a fragment thereof, or as depicted in  FIG. 3A  (SEQ ID NO:9) or a fragment thereof, and wherein said nucleic acid is RNA, the base T (thymine) in SEQ ID NO:7 and SEQ ID NO:8 is substituted by the base U (uracil).  
     
     
         2 . The method of  claim 1 , wherein said cell is contained within an adenoviral or retroviral vector.  
     
     
         3 . The method of  claim 1 , wherein said nucleic acid is a human small cell lung carcinoma cell.  
     
     
         4 . A nutritional formula composition comprising a glycolipid, glycoprotein, glycolipoprotein or oligosaccharide obtained by contacting a recombinant protein comprising an amino acid sequence as depicted in  FIG. 5  (SEQ ID NO:8) with GDP-fucose and a glycolipid, glycoprotein, glycolipoprotein or free oligosaccharide having a terminal Galβ1→3 GalNAc moiety and recovering the glycolipid, glycoprotein, glycolipoprotein, or free oligosaccharide comprising Fucaα1→2Galβ1→3GalNAc.  
     
     
         5 . A nutritional formula composition comprising a glycolipid, glycoprotein, glycolipoprotein or free oligosaccharide comprising Fucα1→2Galβ1→3GalNAc obtained by the method comprising contacting 
 (a) a recombinant rat α1→2fucosyltransferase encoded by the nucleotide sequence depicted as SEQ ID NO: 7, or    (b) a cellular fraction of a recombinant cell containing a vector having and expressing the nucleotide sequence as depicted as SEQ ID NO: 7, said cellular fraction containing the recombinant rat α1→2fucosyltransferase of (a) and having α1→2 fucosyltransferase activity,    with GDP-fucose and a glycolipid, glycoprotein, glycolipoprotein or oligosaccharide having a terminal Galβ1→3GalNAc moiety and recovering a glycolipid, glycoprotein, glycolipoprotein or free oligosaccharide comprising Fucaα1→2Galβ1→3GalNAc and combining to form the nutritional formula composition.    
     
     
         6 . The nutritional formula composition of  claim 4  wherein the α1→2fucosyltransferase is contacted with an oligosaccharide comprising a terminal Galβ1→3GalNAc moiety.  
     
     
         7 . The nutritional formal composition of  claim 5  wherein the rat α1→2fucosyltransferase is contacted with an oligosaccharide comprising a terminal Galβ1→3GalNAc moiety.