ABSTRACT
A bacterial version of human blood group A transferase was identified and found to be able to accept five naturally existing H-antigen core structures as good substrates, demonstrating its versatility for synthesis of blood group A antigens. Furthermore, this enzyme was applied in the engineering of bacterial cell surface polysaccharides by remodeling blood group B mimicry into blood group A mimicry.
Subject(s)
Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Fucosyl Galactose alpha-N-Acetylgalactosaminyltransferase/chemistry , Fucosyl Galactose alpha-N-Acetylgalactosaminyltransferase/metabolism , Helicobacter mustelae/enzymology , ABO Blood-Group System/blood , ABO Blood-Group System/metabolism , Carbohydrate Sequence , Escherichia coli/metabolism , Fucosyl Galactose alpha-N-Acetylgalactosaminyltransferase/blood , Humans , Lipopolysaccharides/chemistry , Lipopolysaccharides/metabolism , Molecular Sequence Data , Substrate SpecificityABSTRACT
We have identified a possible mutation which characterizes A2 alleles (a minor subtype of A) at the human histo-blood group ABO locus based on polymerase chain reaction (PCR) of genomic DNA, followed by nucleotide sequencing of the amplified fragments. The A2 subtype has a single base deletion near the carboxyl terminal. As a result of frame-shifting, A2 transferase possesses an extra domain. Introduction of this single base deletion into the A1 transferase cDNA expression construct drastically decreased the A transferase activity in DNA-transfected HeLa cells.
