Investigation of the nucleotide triphosphate substrate specificity of Homo sapiens UDP-N-acetylgalactosamine pyrophosphorylase (AGX1)

Bioorg Med Chem Lett. 2012 Jun 15;22(12):3957-61. doi: 10.1016/j.bmcl.2012.04.102. Epub 2012 Apr 28.

Abstract

Nucleotide sugars are essential glycosyl donors for Leloir-type glycosyltransferases. The UDP-N-acetylgalactosamine pyrophosphorylase (UDP-GalNAc PP; AGX1) from Homo sapiens catalyzes the synthesis of UDP-N-acetylgalactosamine from N-acetylgalactosamine 1-phosphate and UTP. In this Letter, we systematically studied nucleotide substrate specificity of AGX1 during its uridyltransfer reaction, and described the capability of AGX1 to catalyze dUTP and dTTP to their corresponding nucleotide sugars for the first time. Furthermore, using such a eukaryotic enzyme, we synthesized dUDP-GalNAc and dTDP-GalNAc in multiple mg scale in vitro efficiently and rapidly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylgalactosamine / analogs & derivatives*
  • Acetylgalactosamine / chemistry
  • Biocatalysis
  • Escherichia coli
  • Galactosyltransferases / chemistry*
  • Humans
  • Kinetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Thymine Nucleotides / chemistry*
  • Uridine Diphosphate N-Acetylgalactosamine / chemical synthesis*
  • Uridine Triphosphate / chemistry*

Substances

  • N-acetylgalactosamine-1-phosphate
  • Recombinant Proteins
  • Thymine Nucleotides
  • Uridine Diphosphate N-Acetylgalactosamine
  • Galactosyltransferases
  • UDP-GalNAc pyrophosphorylase AGX1, human
  • Acetylgalactosamine
  • thymidine 5'-triphosphate
  • Uridine Triphosphate