Abstract
Genome sequence data were used to clone and express two sialyltransferase enzymes of the GT-42 family from Helicobacter acinonychis ATCC 51104, a gastric disease isolate from Cheetahs. The deposited genome sequence for these genes contains a large number of tandem repeat sequences in each of them: HAC1267 (RQKELE)(15) and HAC1268 (EEKLLEFKNI)(13). We obtained two clones with different numbers of repeat sequences for the HAC1267 gene homolog and a single clone for the HAC1268 gene homolog. Both genes could be expressed in Escherichia coli and sialyltransferase activity was measured using synthetic acceptor substrates containing a variety of terminal sugars. Both enzymes were shown to have a preference for N-acetyllactosamine, and they each made a product with a different linkage to the terminal galactose. HAC1267 is a mono-functional α2,3-sialyltransferase, whereas HAC1268 is a mono-functional α2,6-sialyltransferase and is the first member of GT-42 to show α2,6-sialyltransferase activity.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Sugars / chemistry
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Bacterial Proteins / biosynthesis
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Carbohydrate Conformation
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Catalytic Domain
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Cloning, Molecular
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Glycosylation
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Helicobacter / enzymology*
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Hydrogen-Ion Concentration
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Magnesium / chemistry
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Manganese / chemistry
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Models, Molecular
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / genetics
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Sialic Acids / chemistry
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Sialyltransferases / biosynthesis
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Sialyltransferases / chemistry*
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Sialyltransferases / genetics
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Structural Homology, Protein
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Substrate Specificity
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beta-D-Galactoside alpha 2-6-Sialyltransferase
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beta-Galactoside alpha-2,3-Sialyltransferase
Substances
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Amino Sugars
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Bacterial Proteins
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Recombinant Fusion Proteins
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Sialic Acids
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N-acetyllactosamine
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Manganese
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Sialyltransferases
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Magnesium
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beta-D-Galactoside alpha 2-6-Sialyltransferase
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beta-Galactoside alpha-2,3-Sialyltransferase