title: The human adenylate kinase 9 is a nucleoside mono- and diphosphate kinase.
abstract: Adenylate kinases regulate adenine nucleotide levels and are present in different intracellular compartments. These enzymes also participate in the activation of pharmacologically active nucleoside and nucleotide analogs. We have in the present study identified the ninth isoform of the adenylate kinase family of enzymes and accordingly named the protein adenylate kinase 9 (AK9). Initially a full-length cDNA of a hypothetical protein containing a predicted adenylate kinase domain was identified and subsequently cloned and expressed in Escherichia coli. The substrate specificity of the recombinant protein showed that the enzyme catalyzed the phosphorylation of AMP, dAMP, CMP and dCMP with ATP as phosphate donor, while only AMP and CMP were phosphorylated when GTP was the phosphate donor. The kinetic parameters of AK9 were determined for AMP, dAMP and CMP with ATP as phosphate donor. Interestingly, in addition to the diphosphate products, a nucleoside diphosphate kinase (NDPK) activity was also present with subsequent triphosphates formed. With ATP or GTP as phosphate donor it was possible to detect the production of ATP, CTP, GTP, UTP, dATP, dCTP, dGTP and TTP as enzymatic products from the corresponding diphosphate substrates. A number of previously characterized adenylate kinases were also tested and found to possess a broad phosphotransferase activity similar to AK9. These enzymes are accordingly suggested to be regarded as nucleoside mono- and diphosphate kinases with catalytic activities possibly determined by local substrate concentrations.
authors: Department of Laboratory Medicine, Division of Clinical Microbiology, Karolinska Institute, F68, SE-141 86 Huddinge, Sweden.
authors: Marjan
authors: M
authors: Amiri
authors: Francesca
authors: F
authors: Conserva
authors: Christakis
authors: C
authors: Panayiotou
authors: Anna
authors: A
authors: Karlsson
authors: Nicola
authors: N
authors: Solaroli
mesh_terms: D000263:Adenylate Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D018929:Cell Culture Techniques; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009703:Nucleoside-Phosphate Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D010957:Plasmids; D013379:Substrate Specificity; D014162:Transfection
chemical_list: D009711:Nucleotides; D000263:Adenylate Kinase; D009703:Nucleoside-Phosphate Kinase; D009701:Nucleoside-Diphosphate Kinase