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Structure of a diguanylate cyclase from Thermotoga maritima: insights into activation, feedback inhibition and thermostability


, : Structure of a diguanylate cyclase from Thermotoga maritima: insights into activation, feedback inhibition and thermostability. Plos One 9(10): E110912-E110912

Large-scale production of bis-3'-5'-cyclic-di-GMP (c-di-GMP) would facilitate biological studies of numerous bacterial signaling pathways and phenotypes controlled by this second messenger molecule, such as virulence and biofilm formation. C-di-GMP constitutes also a potentially interesting molecule as a vaccine adjuvant. Even though chemical synthesis of c-di-GMP can be done, the yields are incompatible with mass-production. tDGC, a stand-alone diguanylate cyclase (DGC or GGDEF domain) from Thermotoga maritima, enables the robust enzymatic production of large quantities of c-di-GMP. To understand the structural correlates of tDGC thermostability, its catalytic mechanism and feedback inhibition, we determined structures of an active-like dimeric conformation with both active (A) sites facing each other and of an inactive dimeric conformation, locked by c-di-GMP bound at the inhibitory (I) site. We also report the structure of a single mutant of tDGC, with the R158A mutation at the I-site, abolishing product inhibition and unproductive dimerization. A comparison with structurally characterized DGC homologues from mesophiles reveals the presence of a higher number of salt bridges in the hyperthermophile enzyme tDGC. Denaturation experiments of mutants disrupting in turn each of the salt bridges unique to tDGC identified three salt-bridges critical to confer thermostability.

US$19.90

PMID: 25360685

DOI: 10.1371/journal.pone.0110912


Other references

Ryall, R.Lyons., 2004: Macromolecules and urolithiasis: parallels and paradoxes. Long-standing interest in the possible role of macromolecules in urolithiasis stems from the observation that all human kidney stones consist of a complex amalgam of mineral and organic material. This review summarises what is currently known or i...

Sung, J.J.Y.; Leung, W.K.; Ling, T.K.W.; Yung, M.Y.; Chan, F.K.L.; Lee, Y.T.; Cheng, A.F.B.; Chung, S.C.S., 1998: One-week use of ranitidine bismuth citrate, amoxycillin and clarithromycin for the treatment of Helicobacter pylori-related duodenal ulcer. Background: Proton pump inhibitors have been widely used in combination with amoxycillin, clarithromycin or metronidazole for the treatment of Helicobacter pylori infection. Aim: To study the effects of 1-week ranitidine bismuth citrate (RBC)-base...

Kunwar, A.; Barik, A.; Mishra, B.; Rathinasamy, K.; Pandey, R.; Priyadarsini, K.I., 2008: Quantitative cellular uptake, localization and cytotoxicity of curcumin in normal and tumor cells. Using absorption and fluorescence spectroscopic methods, quantitative cellular uptake of curcumin, an antioxidant and anti-tumor agent from Curcuma longa, was calculated in two types of normal cells: spleen lymphocytes, and NIH3T3 and two tumor c...

Petereit, H.F.; Richter, N.; Pukrop, R.; Bamborschke, S., 2000: Interferon gamma production in blood lymphocytes correlates with disability score in multiple sclerosis patients. The proinflammatory cytokine interferon gamma (IFG) is elevated in body fluids of multiple sclerosis patients but its variation range is broad. The reason for this wide scatter of IFG production is not yet known. We looked for the relation between...

Bergman, E.L.; Autio, K.; Sandberg, A.S., 2000: Optimal conditions for phytate degradation, estimation of phytase activity, and localization of phytate in barley (cv. Blenheim). Using a multivariate experimental design, optimal conditions for phytate degradation were found to be pH 4.8 and 57 degrees C in barley flour (cv. Blenheim) and pH 5.2 and 47 degrees C in a crude extracted phytase from barley. Three methods for me...

Mathias Mildred E., 1965: Distribution patterns of certain Umbelliferae. Modern world distribution patterns of the angiosperm family Umbelliferae and the closely related, less advanced Araliaceae are plotted from compiled data, and Paleocene-Pleistocene occurrences of species in North America are tabulated. The two fam...

Bleeker G.M., 1982: Injuries to the eye sockets. Klinische Monatsblaetter Fuer: 246

Sharma, S.N.; Nilakanitan, P.R., 1964: Observations on the haemolytic activity of Newcastle (Ranikhet) disease virus. The haemolytic activity of five strains produced in amnio-allantoic fluids was studied. A two-fold variation in susceptibility of r.b.c. from 10 individual fowls was noted. Virus dilutions were used untreated or after repeated freezing and thawing...

Grabetskaya N.A., 1990: Gravity differentiation as a criterion of reconstruction of sedimentation conditions. Byulleten' Komissiy po Izucheniyu Chetvertichnogo Perioda 59: 58-65

Berkley, K.J.; Miles, J.B.; Bowsher, D.R., 2012: David R. Bowsher, M.D., Sc.D., Ph.D., FRCPEd., FRCPath., 1925–2011. Pain 152(10): 2189-2190