Publications

• The biosynthesis of trehalose phosphate.

  CABIB E., LELOIR L.F.

  J Biol Chem 231:259-275(1958) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Mechanistic insight into enzymatic glycosyl transfer with retention of configuration through analysis of glycomimetic inhibitors.

  Errey J.C., Lee S.S., Gibson R.P., Martinez Fleites C., Barry C.S., Jung P.M., O'Sullivan A.C., Davis B.G., Davies G.J.

  Angew. Chem. Int. Ed. Engl. 49:1234-1237(2010) [PubMed] [EuropePMC]

• Characterization of trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase of Saccharomyces cerevisiae.

  Vandercammen A., Francois J., Hers H.-G.

  The properties of yeast trehalose-6-phosphate synthase were reinvestigated in relation with the recent claim made by Panek et al. [Panek, A. C., de Araujo, P. S., Moura-Neto, V. and Panek, A. D. (1987) Curr. Genet. II, 459-465] that the enzyme would be stimulated by ATP and partially inactivated b ... >> More

  The properties of yeast trehalose-6-phosphate synthase were reinvestigated in relation with the recent claim made by Panek et al. [Panek, A. C., de Araujo, P. S., Moura-Neto, V. and Panek, A. D. (1987) Curr. Genet. II, 459-465] that the enzyme would be stimulated by ATP and partially inactivated by cAMP-dependent protein kinase. Trehalose-6-phosphate synthase activity was measured by the sum of [14C]trehalose 6-phosphate and [14C]trehalose formed from UDP-[14C]glucose and glucose 6-phosphate. The activity measured in an extract of Saccharomyces cerevisiae was not affected by any treatment of the cells, such as incubation in the presence of glucose or of dinitrophenol, which are known to greatly increase the intracellular concentration of cAMP, nor by preincubation of the extract in the presence of ATP-Mg, cAMP and bovine heart cAMP-dependent protein kinase. The activity was also not significantly different in several mutants affected in the cAMP system. The kinetic properties of the partially purified enzyme were investigated; no effect of ATP could be detected but Pi acted as a potent noncompetitive inhibitor (Ki = 2 mM). The activity of trehalose-6-phosphate phosphatase was measured by the amount of [14C]trehalose formed from [14C]trehalose 6-phosphate. The enzyme could be separated from other phosphatases and appeared to be highly specific for trehalose 6-phosphate. It was Mg dependent and its kinetics for trehalose 6-phosphate was hyperbolic. Studies performed with intact cells, crude extracts or the purified enzyme did not reveal any cAMP-dependent change in its activity. Remarkably, trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase copurified in the course of different chromatographic procedures, suggesting that they are part of a single bifunctional protein. A 50-fold purification of the two enzymes could be achieved with a yield of only 2% by chromatography on Mono S followed by gel filtration on Superose 6B. << Less

  Eur. J. Biochem. 182:613-620(1989) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• The first prokaryotic trehalose synthase complex identified in the hyperthermophilic crenarchaeon Thermoproteus tenax.

  Zaparty M., Hagemann A., Brasen C., Hensel R., Lupas A.N., Brinkmann H., Siebers B.

  The role of the disaccharide trehalose, its biosynthesis pathways and their regulation in Archaea are still ambiguous. In Thermoproteus tenax a fused trehalose-6-phosphate synthase/phosphatase (TPSP), consisting of an N-terminal trehalose-6-phosphate synthase (TPS) and a C-terminal trehalose-6-pho ... >> More

  The role of the disaccharide trehalose, its biosynthesis pathways and their regulation in Archaea are still ambiguous. In Thermoproteus tenax a fused trehalose-6-phosphate synthase/phosphatase (TPSP), consisting of an N-terminal trehalose-6-phosphate synthase (TPS) and a C-terminal trehalose-6-phosphate phosphatase (TPP) domain, was identified. The tpsp gene is organized in an operon with a putative glycosyltransferase (GT) and a putative mechanosensitive channel (MSC). The T. tenax TPSP exhibits high phosphatase activity, but requires activation by the co-expressed GT for bifunctional synthase-phosphatase activity. The GT mediated activation of TPS activity relies on the fusion of both, TPS and TPP domain, in the TPSP enzyme. Activation is mediated by complex-formation in vivo as indicated by yeast two-hybrid and crude extract analysis. In combination with first evidence for MSC activity the results suggest a sophisticated stress response involving TPSP, GT and MSC in T. tenax and probably in other Thermoproteales species. The monophyletic prokaryotic TPSP proteins likely originated via a single fusion event in the Bacteroidetes with subsequent horizontal gene transfers to other Bacteria and Archaea. Furthermore, evidence for the origin of eukaryotic TPSP fusions via HGT from prokaryotes and therefore a monophyletic origin of eukaryotic and prokaryotic fused TPSPs is presented. This is the first report of a prokaryotic, archaeal trehalose synthase complex exhibiting a much more simple composition than the eukaryotic complex described in yeast. Thus, complex formation and a complex-associated regulatory potential might represent a more general feature of trehalose synthesizing proteins. << Less

  PLoS ONE 8:E61354-E61354(2013) [PubMed] [EuropePMC]

• PURIFICATION AND PROPERTIES OF THE TRANSGLUCOSYLASE INHIBITOR OF MYCOBACTERIUM TUBERCULOSIS.

  LORNITZO F.A., GOLDMAN D.S.

  J Biol Chem 239:2730-2734(1964) [PubMed] [EuropePMC]

• Biochemical and genetic characterization of osmoregulatory trehalose synthesis in Escherichia coli.

  Giaever H.M., Styrvold O.B., Kaasen I., Stroem A.R.

  It has been shown previously that Escherichia coli accumulates endogenously synthesized trehalose under osmotic stress. We report here that E. coli contained an osmotically regulated trehalose-phosphate synthase which utilized UDP-glucose and glucose 6-phosphate as substrates. In the wild type, th ... >> More

  It has been shown previously that Escherichia coli accumulates endogenously synthesized trehalose under osmotic stress. We report here that E. coli contained an osmotically regulated trehalose-phosphate synthase which utilized UDP-glucose and glucose 6-phosphate as substrates. In the wild type, the synthase was induced by growth in glucose-mineral medium of elevated osmotic strength and the synthase itself was strongly stimulated by K+ and other monovalent cations. A laboratory strain which expressed the synthase at a high constitutive level was found. GalU mutants, defective in synthesis of UDP-glucose, did not accumulate trehalose. Two genes governing the synthase were identified and named otsA and otsB (osmoregulatory trehalose synthesis). They mapped near 42 min in the flbB-uvrC region. Mutants with an otsA-lacZ or otsB-lacZ operon fusion displayed osmotically inducible beta-galactosidase activity; i.e., the activity was increased fivefold by growth in medium of elevated osmotic strength. Mutants unable to synthesize trehalose (galU, otsA, and otsB) were osmotically sensitive in glucose-mineral medium. But an osmotically tolerant phenotype was restored in the presence of glycine betaine, which also partially repressed the synthesis of synthase in the wild type and of beta-galactosidase in ots-lacZ fusion mutants. << Less

  J. Bacteriol. 170:2841-2849(1988) [PubMed] [EuropePMC]

• Trehalose-phosphate synthase of Mycobacterium tuberculosis. Cloning, expression and properties of the recombinant enzyme.

  Pan Y.T., Carroll J.D., Elbein A.D.

  The trehalose-phosphate synthase (TPS) of Mycobacterium smegmatis was previously purified to apparent homogeneity and several peptides from the 58 kDa protein were sequenced. Based on that sequence information, the gene for TPS was identified in the Mycobacterium tuberculosis genome, and the gene ... >> More

  The trehalose-phosphate synthase (TPS) of Mycobacterium smegmatis was previously purified to apparent homogeneity and several peptides from the 58 kDa protein were sequenced. Based on that sequence information, the gene for TPS was identified in the Mycobacterium tuberculosis genome, and the gene was cloned and expressed in Escherichia coli with a (His)6 tag at the amino terminus. The TPS was expressed in good yield and as active enzyme, and was purified on a metal ion column to give a single band of approximately 58 kDa on SDS/PAGE. Approximately 1.3 mg of purified TPS were obtained from a 1-L culture of E. coli ( approximately 2.3 g cell paste). The purified recombinant enzyme showed a single band of approximately 58 kDa on SDS/PAGE, but a molecular mass of approximately 220 kDa by gel filtration, indicating that the active TPS is probably a tetrameric protein. Like the enzyme originally purified from M. smegmatis, the recombinant enzyme is an unusual glycosyltransferase as it can utilize any of the nucleoside diphosphate glucose derivatives as glucosyl donors, i.e. ADP-glucose, CDP-glucose, GDP-glucose, TDP-glucose and UDP-glucose, with ADP-glucose, GDP-glucose and UDP-glucose being the preferred substrates. These studies prove conclusively that the mycobacterial TPS is indeed responsible for catalyzing the synthesis of trehalose-P from any of the nucleoside diphosphate glucose derivatives. Although the original enzyme from M. smegmatis was greatly stimulated in its utilization of UDP-glucose by polyanions such as heparin, the recombinant enzyme was stimulated only modestly by heparin. The Km for UDP-glucose as the glucosyl donor was approximately 18 mm, and that for GDP-glucose was approximately 16 mm. The enzyme was specific for glucose-6-P as the glucosyl acceptor, and the Km for this substrate was approximately 7 mm when UDP-glucose was the glucosyl donor and approximately 4 mm with GDP-glucose. TPS did not show an absolute requirement for divalent cations, but activity was increased about twofold by 10 mm Mn2+. This recombinant system will be useful for obtaining sufficient amounts of protein for structural studies. TPS should be a valuable target site for chemotherapeutic intervention in tuberculosis. << Less

  Eur. J. Biochem. 269:6091-6100(2002) [PubMed] [EuropePMC]

  This publication is cited by 5 other entries.

• Structures of trehalose-6-phosphate synthase, Tps1, from the fungal pathogen <i>Cryptococcus neoformans</i>: A target for antifungals.

  Washington E.J., Zhou Y., Hsu A.L., Petrovich M., Tenor J.L., Toffaletti D.L., Guan Z., Perfect J.R., Borgnia M.J., Bartesaghi A., Brennan R.G.

  Invasive fungal diseases are a major threat to human health, resulting in more than 1.5 million annual deaths worldwide. The arsenal of antifungal therapeutics remains limited and is in dire need of drugs that target additional biosynthetic pathways that are absent from humans. One such pathway in ... >> More

  Invasive fungal diseases are a major threat to human health, resulting in more than 1.5 million annual deaths worldwide. The arsenal of antifungal therapeutics remains limited and is in dire need of drugs that target additional biosynthetic pathways that are absent from humans. One such pathway involves the biosynthesis of trehalose. Trehalose is a disaccharide that is required for pathogenic fungi to survive in their human hosts. In the first step of trehalose biosynthesis, trehalose-6-phosphate synthase (Tps1) converts UDP-glucose and glucose-6-phosphate to trehalose-6-phosphate. Here, we report the structures of full-length <i>Cryptococcus neoformans</i> Tps1 (CnTps1) in unliganded form and in complex with uridine diphosphate and glucose-6-phosphate. Comparison of these two structures reveals significant movement toward the catalytic pocket by the N terminus upon ligand binding and identifies residues required for substrate binding, as well as residues that stabilize the tetramer. Intriguingly, an intrinsically disordered domain (IDD), which is conserved among Cryptococcal species and closely related basidiomycetes, extends from each subunit of the tetramer into the "solvent" but is not visible in density maps. We determined that the IDD is not required for <i>C. neoformans</i> Tps1-dependent thermotolerance and osmotic stress survival. Studies with UDP-galactose highlight the exquisite substrate specificity of CnTps1. In toto, these studies expand our knowledge of trehalose biosynthesis in <i>Cryptococcus</i> and highlight the potential of developing antifungal therapeutics that disrupt the synthesis of this disaccharide or the formation of a functional tetramer and the use of cryo-EM in the structural characterization of CnTps1-ligand/drug complexes. << Less

  Proc Natl Acad Sci U S A 121:e2314087121-e2314087121(2024) [PubMed] [EuropePMC]

• THE ENZYMES OF GLYCOGEN AND TREHALOSE SYNTHESIS IN SILK MOTH FAT BODY.

  MURPHY T.A., WYATT G.R.

  J Biol Chem 240:1500-1508(1965) [PubMed] [EuropePMC]

• The biosynthesis of trehalose in the locust fat body.

  CANDY D.J., KILBY B.A.

  Biochem J 78:531-536(1961) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.