Characterization of α2,3- and α2,6-sialyltransferases from Helicobacter acinonychis

Glycobiology. 2012 Jul;22(7):997-1006. doi: 10.1093/glycob/cws071. Epub 2012 Apr 14.

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

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Sugars / chemistry
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Carbohydrate Conformation
  • Catalytic Domain
  • Cloning, Molecular
  • Glycosylation
  • Helicobacter / enzymology*
  • Hydrogen-Ion Concentration
  • Magnesium / chemistry
  • Manganese / chemistry
  • Models, Molecular
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / genetics
  • Sialic Acids / chemistry
  • Sialyltransferases / biosynthesis
  • Sialyltransferases / chemistry*
  • Sialyltransferases / genetics
  • Structural Homology, Protein
  • Substrate Specificity
  • beta-D-Galactoside alpha 2-6-Sialyltransferase
  • beta-Galactoside alpha-2,3-Sialyltransferase

Substances

  • Amino Sugars
  • Bacterial Proteins
  • Recombinant Fusion Proteins
  • Sialic Acids
  • N-acetyllactosamine
  • Manganese
  • Sialyltransferases
  • Magnesium
  • beta-D-Galactoside alpha 2-6-Sialyltransferase
  • beta-Galactoside alpha-2,3-Sialyltransferase