Phosphorylation of 9-beta-D-arabinofuranosylguanine monophosphate by Drosophila melanogaster guanylate kinase

Biochem Pharmacol. 2005 Oct 1;70(7):987-92. doi: 10.1016/j.bcp.2005.06.023.

Abstract

Nucleoside monophosphate kinases have an important role in the synthesis of nucleotides that are required for cellular metabolism. These enzymes are also important for the phosphorylation of nucleoside- and nucleotide analogs used in cancer and anti-viral therapy. We report the cDNA cloning and characterization of a 23 kDa guanylate kinase from Drosophila melanogaster (Dm-GUK). The predicted amino acid sequence was 58% identical to the human guanylate kinase and the enzyme was shown to phosphorylate GMP and dGMP with ATP as phosphate donor. The monophosphates of the deoxyguanosine analogs 2',2'-difluorodeoxyguanosine (dFdG) and 9-beta-D-arabinofuranosylguanine (araG) were also shown to be phosphorylated by the enzyme. We used the enzyme to reconstitute the complete in vitro three-step phosphorylation pathway for the conversion of dGuo and araG to the corresponding triphosphates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arabinonucleotides / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • DNA, Complementary
  • Drosophila melanogaster / enzymology*
  • Guanylate Kinases
  • Molecular Sequence Data
  • Nucleoside-Phosphate Kinase / chemistry
  • Nucleoside-Phosphate Kinase / genetics
  • Nucleoside-Phosphate Kinase / metabolism*
  • Phosphorylation
  • Sequence Homology, Amino Acid
  • Substrate Specificity

Substances

  • 9-beta-D-arabinofuranosylguanine monophosphate
  • Arabinonucleotides
  • DNA Primers
  • DNA, Complementary
  • Nucleoside-Phosphate Kinase
  • Guanylate Kinases