Sugar-nucleotide donor specificity of histo-blood group A and B transferases is based on amino acid substitutions.
Four amino acid substitutions (aa 176, 235, 266, and 268) have been found between the coding regions of cDNAs for histo-blood group A and B transferases (Yamamoto F., Clausen, H., White, T., Marken, J., and Hakomori, S. (1990) Nature 345, 229-233). Here we establish the basis of differential affinity of these glycosyltransferases to nucleotide-sugar (UDP-GalNAc or UDP-Gal). On the basis of gene reconstruction experiments and studies of expression in DNA transfected HeLa cells, the third as well as the fourth aa substitutions (leucine and glycine in A and methionine and alanine in B), which were calculated to modify flexibility of the protein, were found to be crucial in determining nucleotide-sugar specificity. The second substitution (glycine in A and serine in B) also affects the specificity. We have also created new enzymes which catalyze the transfer of both GalNAc and Gal, and may provide an explanation of the rare cis-AB phenotype.
Yamamoto
F
F
Department of Molecular Biology, Biomembrane Institute, Seattle, Washington 98119.
Hakomori
S
S
D000017:ABO Blood-Group System / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002678:Chimera; D003001:Cloning, Molecular; D005700:Galactosyltransferases / Q000235:genetics*; D005838:Genotype; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D017350:N-Acetylgalactosaminyltransferases; D015345:Oligonucleotide Probes; D010641:Phenotype; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D000017:ABO Blood-Group System; D015345:Oligonucleotide Probes; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C003742:UDPgalactosamine-galactose acetylgalactosaminyltransferase; C023155:blood-group-substance alpha-D-galactosyltransferase