title: Phosphorylation of 9-beta-D-arabinofuranosylguanine monophosphate by Drosophila melanogaster guanylate kinase.
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.
authors.affiliation: Karolinska Institute, Department of Laboratory Medicine, Novum, S-141 57 Stockholm, Sweden.
authors.forename: Magnus
authors.initials: M
authors.lastname: Johansson
authors.forename: Marjan
authors.initials: M
authors.lastname: Amiri
authors.forename: Anna
authors.initials: A
authors.lastname: Karlsson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001088:Arabinonucleotides / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004331:Drosophila melanogaster / Q000201:enzymology*; D051528:Guanylate Kinases; D008969:Molecular Sequence Data; D009703:Nucleoside-Phosphate Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: C505040:9-beta-D-arabinofuranosylguanine monophosphate; D001088:Arabinonucleotides; D017931:DNA Primers; D018076:DNA, Complementary; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases