Publications

• Dihydroxyacetone metabolism by human erythrocytes: demonstration of triokinase activity and its characterization.

  Beutler E., Guinto E.

  Blood 41:559-568(1973) [PubMed] [EuropePMC]

  This publication is cited by 2 other entries.

• Crystal structure of the Citrobacter freundii dihydroxyacetone kinase reveals an eight-stranded alpha-helical barrel ATP-binding domain.

  Siebold C., Arnold I., Garcia-Alles L.F., Baumann U., Erni B.

  Dihydroxyacetone kinases are a sequence-conserved family of enzymes, which utilize two different phosphoryldonors, ATP in animals, plants and some bacteria, and a multiphosphoprotein of the phosphoenolpyruvate carbohydrate phosphotransferase system in bacteria. Here we report the 2.5-A crystal str ... >> More

  Dihydroxyacetone kinases are a sequence-conserved family of enzymes, which utilize two different phosphoryldonors, ATP in animals, plants and some bacteria, and a multiphosphoprotein of the phosphoenolpyruvate carbohydrate phosphotransferase system in bacteria. Here we report the 2.5-A crystal structure of the homodimeric Citrobacter freundii dihydroxyacetone kinase complex with an ATP analogue and dihydroxyacetone. The N-terminal domain consists of two alpha/beta-folds with a molecule of dihydroxyacetone covalently bound in hemiaminal linkage to the N epsilon 2 of His-220. The C-terminal domain consists of a regular eight-helix alpha-barrel. The eight helices form a deep pocket, which includes a tightly bound phospholipid. Only the lipid headgroup protrudes from the surface. The nucleotide is bound on the top of the barrel across from the entrance to the lipid pocket. The phosphate groups are coordinated by two Mg2+ ions to gamma-carboxyl groups of aspartyl residues. The ATP binding site does not contain positively charged or aromatic groups. Paralogues of dihydroxyacetone kinase also occur in association with transcription regulators and proteins of unknown function pointing to biological roles beyond triose metabolism. << Less

  J. Biol. Chem. 278:48236-48244(2003) [PubMed] [EuropePMC]

• Identification of human and rat FAD-AMP lyase (cyclic FMN forming) as ATP-dependent dihydroxyacetone kinases.

  Cabezas A., Costas M.J., Pinto R.M., Couto A., Cameselle J.C.

  Rat liver FAD-AMP lyase or FMN cyclase is the only known enzymatic source of the unusual flavin nucleotide riboflavin 4',5'-cyclic phosphate. To determine its molecular identity, a peptide-mass fingerprint of the purified rat enzyme was obtained. It pointed to highly related, mammalian hypothetica ... >> More

  Rat liver FAD-AMP lyase or FMN cyclase is the only known enzymatic source of the unusual flavin nucleotide riboflavin 4',5'-cyclic phosphate. To determine its molecular identity, a peptide-mass fingerprint of the purified rat enzyme was obtained. It pointed to highly related, mammalian hypothetical proteins putatively classified as dihydroxyacetone (Dha) kinases due to weaker homologies to biochemically proven Dha kinases of plants, yeasts, and bacteria. The human protein LOC26007 cDNA was used to design PCR primers. The product amplified from human brain cDNA was cloned, sequenced (GenBank Accession No. ), and found to differ from protein LOC26007 cDNA by three SNPs. Its heterologous expression yielded a protein active both as FMN cyclase and ATP-dependent Dha kinase, each activity being inhibited by the substrate(s) of the other. Cyclase and kinase activities copurified from rat liver extracts. Evidence supports that a single protein sustains both activities, probably in a single active center. Putative Dha kinases from other mammals are likely to be FMN cyclases too. Future work will profit from the availability of the structure of Citrobacter freundii Dha kinase, which contains substrate-interacting residues conserved in human Dha kinase/FMN cyclase. << Less

  Biochem. Biophys. Res. Commun. 338:1682-1689(2005) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Biochemical and molecular characterization of the oxidative branch of glycerol utilization by Citrobacter freundii.

  Daniel R., Stuertz K., Gottschalk G.

  Glycerol dehydrogenase (EC 1.1.1.6) and dihydroxyacetone kinase (EC 2.7.1.29) were purified from Citrobacter freundii. The dehydrogenase is a hexamer of a polypeptide of 43,000 Da. The enzyme exhibited a rather broad substrate specificity, but glycerol was the preferred substrate in the physiologi ... >> More

  Glycerol dehydrogenase (EC 1.1.1.6) and dihydroxyacetone kinase (EC 2.7.1.29) were purified from Citrobacter freundii. The dehydrogenase is a hexamer of a polypeptide of 43,000 Da. The enzyme exhibited a rather broad substrate specificity, but glycerol was the preferred substrate in the physiological direction. The apparent Kms of the enzyme for glycerol and NAD+ were 1.27 mM and 57 microM, respectively. The kinase is a dimer of a polypeptide of 57,000 Da. The enzyme was highly specific for the substrates dihydroxyacetone and ATP; the apparent Kms were 30 and 70 microM, respectively. The DNA region which contained the genes encoding glycerol dehydrogenase (dhaD) and dihydroxyacetone kinase (dhaK) was cloned and sequenced. Both genes were identified by N-terminal sequence comparison. The deduced dhaD gene product (365 amino acids) exhibited high degrees of homology to glycerol dehydrogenases from other organisms and less homology to type III alcohol dehydrogenases, whereas the dhaK gene product (552 amino acids) revealed no significant homology to any other protein in the databases. A large gene (dhaR) of 1,929 bp was found downstream from dhaD. The deduced gene product (641 amino acids) showed significant similarities to members of the sigma 54 bacterial enhancer-binding protein family. << Less

  J. Bacteriol. 177:4392-4401(1995) [PubMed] [EuropePMC]

• Purification and characterization of triokinase from porcine kidney.

  Miwa I., Kito Y., Okuda J.

  In order to be able to use triokinase for the enzymatic assay of tissue glyceraldehyde, we purified the enzyme to homogeneity from porcine kidney and characterized its biochemical properties. The purification was performed by polyethylene glycol fractionation, anion exchange chromatography, hydrox ... >> More

  In order to be able to use triokinase for the enzymatic assay of tissue glyceraldehyde, we purified the enzyme to homogeneity from porcine kidney and characterized its biochemical properties. The purification was performed by polyethylene glycol fractionation, anion exchange chromatography, hydroxyapatite chromatography, hydrophobic chromatography, and gel filtration. The enzyme was purified 937-fold from the crude extract with an overall yield of 28%. It had a molecular weight of 122,000 and was a dimer composed of identical subunits. The optimal pH and optimal temperature were 7.0 and 60 degrees C, respectively. This enzyme was stable when incubated at pH 7.0 at 40 degrees C for 1 h in the presence of 0.1 mg/ml bovine serum albumin. No loss of activity occurred for at least 1 month when the enzyme was stored at 4 degrees C in the presence of 1 mM dithiothreitol and 15 mM NaN3 under N2. Only three compounds, i.e., D-glyceraldehyde, dihydroxyacetone, and glycolaldehyde, acted as the substrate of the enzyme, having Km's of 11, < 5, and 260 microM, respectively. The Km for ATP-Mg2+ was 68 microM. These results indicate that porcine kidney triokinase has properties advantageous for the glyceraldehyde assay using glyceraldehyde-3-phosphate dehydrogenase as a coupling enzyme. << Less

  Prep. Biochem. 24:203-223(1994) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.