Protein backbone stability resonance free#

Golovanov AP, Hautbergue GM, Wilson SA, Lian L-Y (2004) A simple method for improving protein solubility and long-term stability. George GM, van der Merwe MJ, Nunes-Nesi A, Bauer R, Fernie AR, Kossmann J, Lloyd JR (2010) Virus-induced gene silencing of plastidial soluble inorganic pyrophosphatase impairs essential leaf anabolic pathways and reduces drought stress tolerance in Nicotiana benthamiana. Galizzi M, Bustamante JM, Fang J, Miranda K, Soares Medeiros LC, Tarleton RL, Docampo R (2013) Evidence for the role of vacuolar soluble pyrophosphatase and inorganic polyphosphate in Trypanosoma cruzi persistence.

Protein backbone stability resonance free#

J Biomol NMR 6(3):277–293įarrow NA, Muhandiram R, Singer AU, Pascal SM, Kay CM, Gish G, Shoelson SE, Pawson T, Forman-Kay JD, Kay LE (1994) Backbone dynamics of a free and phosphopeptide-complexed Src homology 2 domain studied by 15N NMR relaxation. Trends Biochem Sci 17(7):262–266ĭelaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. Ĭooperman BS, Baykov AA, Lahti R (1992) Evolutionary conservation of the active site of soluble inorganic pyrophosphatase. Springer, Berlin, pp 127–150Ĭhen J, Brevet A, Fromant M, Lévêque F, Schmitter JM, Blanquet S, Plateau P (1990) Pyrophosphatase is essential for growth of Escherichia coli.

In: Schröder HC, Muller WEG (eds) Inorganic polyphosphates. PLoS Comput Biol 5(3):e1000307īaykov A, Cooperman B, Goldman A, Lahti R (1999) Cytoplasmic inorganic pyrophosphatase. īahrami A, Assadi AH, Markley JL, Eghbalnia HR (2009) Probabilistic interaction network of evidence algorithm and its application to complete labeling of peak lists from protein NMR spectroscopy. Infect Immun 66(1):203–212Īlcock NW, Shils ME (1969) Association of inorganic pyrophosphatase activity with normal calcification of rat costal cartilage in vivo. Determined resonance assignments and dynamic properties provide the basis for the subsequent structure-based design of novel inhibitors of Mt-PPase-potential anti-tuberculosis drugs.Ību Kwaik Y (1998) Induced expression of the Legionella pneumophila gene encoding a 20-kilodalton protein during intracellular infection. Protein backbone dynamics were studied using 15N NMR relaxation experiments. It was found that the secondary structure of Mt-PPase in solution corresponds well to the crystal structure of this protein. Backbone resonance assignment was achieved for more than 95% of the residues. Samples of Mt-PPase enriched with 99% of 13C and 15N isotopes, and 95% of 2H were obtained using recombinant protein expression in an isotopically-labeled medium and effective heat-shock protocol for the deuterium-to-hydrogen exchange of the amide groups. In order to use NMR techniques in functional studies of Mt-PPase and rational design of the inhibitors for this enzyme, it is necessary to have information on the backbone 1H, 13C and 15N resonance assignments. Hexameric 110 kDa (6 × 18.3 kDa) PPase from Mycobacterium tuberculosis (Mt-PPase) is a promising target for the rational design of potential anti-tuberculosis agents.

PPases from various organisms are extensively studied by X-ray crystallography but until now there was no information on their structure and dynamics in solution.

Soluble PPases retain enzymatic activity only in multimeric forms. They hydrolyze inorganic pyrophosphate, thus providing the driving force for numerous biosynthetic reactions. Family I soluble inorganic pyrophosphatases (PPases EC 3.6.1.1) are enzymes essential for all organisms.