Publications

• An extremely thermostable aldolase from Sulfolobus solfataricus with specificity for non-phosphorylated substrates.

  Buchanan C.L., Connaris H., Danson M.J., Reeve C.D., Hough D.W.

  Sulfolobus solfataricus is a hyperthermophilic archaeon growing optimally at 80-85 degrees C. It metabolizes glucose via a novel non-phosphorylated Entner-Doudoroff pathway, in which the reversible C(6) to C(3) aldol cleavage is catalysed by 2-keto-3-deoxygluconate aldolase (KDG-aldolase), generat ... >> More

  Sulfolobus solfataricus is a hyperthermophilic archaeon growing optimally at 80-85 degrees C. It metabolizes glucose via a novel non-phosphorylated Entner-Doudoroff pathway, in which the reversible C(6) to C(3) aldol cleavage is catalysed by 2-keto-3-deoxygluconate aldolase (KDG-aldolase), generating pyruvate and glyceraldehyde. Given the ability of such a hyperstable enzyme to catalyse carbon-carbon-bond synthesis with non-phosphorylated metabolites, we report here the cloning and sequencing of the S. solfataricus gene encoding KDG-aldolase, and its expression in Escherichia coli to give fully active enzyme. The recombinant enzyme was purified in a simple two-step procedure, and shown to possess kinetic properties indistinguishable from the enzyme purified from S. solfataricus cells. The KDG-aldolase is a thermostable tetrameric protein with a half-life at 100 degrees C of 2.5 h, and is equally active with both d- and l-glyceraldehyde. It exhibits sequence similarity to the N-acetylneuraminate lyase superfamily of Schiff-base-dependent aldolases, dehydratases and decarboxylases, and evidence is presented for a similar catalytic mechanism for the archaeal enzyme by substrate-dependent inactivation by reduction with NaBH(4). << Less

  Biochem. J. 343:563-570(1999) [PubMed] [EuropePMC]

• Prediction of enzymatic pathways by integrative pathway mapping.

  Calhoun S., Korczynska M., Wichelecki D.J., San Francisco B., Zhao S., Rodionov D.A., Vetting M.W., Al-Obaidi N.F., Lin H., O'Meara M.J., Scott D.A., Morris J.H., Russel D., Almo S.C., Osterman A.L., Gerlt J.A., Jacobson M.P., Shoichet B.K., Sali A.

  The functions of most proteins are yet to be determined. The function of an enzyme is often defined by its interacting partners, including its substrate and product, and its role in larger metabolic networks. Here, we describe a computational method that predicts the functions of orphan enzymes by ... >> More

  The functions of most proteins are yet to be determined. The function of an enzyme is often defined by its interacting partners, including its substrate and product, and its role in larger metabolic networks. Here, we describe a computational method that predicts the functions of orphan enzymes by organizing them into a linear metabolic pathway. Given candidate enzyme and metabolite pathway members, this aim is achieved by finding those pathways that satisfy structural and network restraints implied by varied input information, including that from virtual screening, chemoinformatics, genomic context analysis, and ligand -binding experiments. We demonstrate this integrative pathway mapping method by predicting the L-gulonate catabolic pathway in <i>Haemophilus influenzae</i> Rd KW20. The prediction was subsequently validated experimentally by enzymology, crystallography, and metabolomics. Integrative pathway mapping by satisfaction of structural and network restraints is extensible to molecular networks in general and thus formally bridges the gap between structural biology and systems biology. << Less

  Elife 7:e31097-e31097(2018) [PubMed] [EuropePMC]

  This publication is cited by 4 other entries.

• CRYSTALLIZATION AND CHARACTERISTICS OF 2-KETO-3-DEOXY-6-PHOSPHOGLUCONIC ALDOLASE.

  MELOCHE H.P., WOOD W.A.

  J Biol Chem 239:3515-3518(1964) [PubMed] [EuropePMC]

• Promiscuity in the part-phosphorylative Entner-Doudoroff pathway of the archaeon Sulfolobus solfataricus.

  Lamble H.J., Theodossis A., Milburn C.C., Taylor G.L., Bull S.D., Hough D.W., Danson M.J.

  The hyperthermophilic archaeon Sulfolobus solfataricus metabolises glucose and galactose by a 'promiscuous' non-phosphorylative variant of the Entner-Doudoroff pathway, in which a series of enzymes have sufficient substrate promiscuity to permit the metabolism of both sugars. Recently, it has been ... >> More

  The hyperthermophilic archaeon Sulfolobus solfataricus metabolises glucose and galactose by a 'promiscuous' non-phosphorylative variant of the Entner-Doudoroff pathway, in which a series of enzymes have sufficient substrate promiscuity to permit the metabolism of both sugars. Recently, it has been proposed that the part-phosphorylative Entner-Doudoroff pathway occurs in parallel in S. solfataricus as an alternative route for glucose metabolism. In this report we demonstrate, by in vitro kinetic studies of D-2-keto-3-deoxygluconate (KDG) kinase and KDG aldolase, that the part-phosphorylative pathway in S. solfataricus is also promiscuous for the metabolism of both glucose and galactose. << Less

  FEBS Lett. 579:6865-6869(2005) [PubMed] [EuropePMC]

  This publication is cited by 2 other entries.

• The mechanism of 2-keto-3-deoxy-6-phosphogluconate aldolase. 3. Nature of the inactivation by fluorodinitrobenzene.

  Barran L.R., Wood W.A.

  J Biol Chem 246:4028-4035(1971) [PubMed] [EuropePMC]