Plasmodium falciparum dUTPase: Studies on protein stability and binding of deoxyuridine derivatives

Quesada-Soriano, Indalecio; Musso-Buendia, Juan Alexander; Téllez-Sanz, Ramiro; Ruíz-Pérez, Luis M.; Barón, Carmen; González-Pacanowska, Dolores; Garcı́a-Fuentes, Luis

doi:10.1016/j.bbapap.2007.04.013

Cited by 11 publications

(4 citation statements)

References 36 publications

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“…In order to check folding and stability of the recombinant protein, differential scanning calorimetry was applied [11,21,22]. Melting temperature was found to be 61.7 °C, closely similar to the value obtained for Drosophila melanogaster or Plasmodium falciparum dUTPases (56.5 °C and 63.8 °C, respectively) and considerably lower than for E. coli dUTPase (74.7 °C), indicating decreased stability of eukaryotic dUTPases [21,22].…”

Section: Resultssupporting

confidence: 65%

Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase

et al. 2007

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Human dUTPase, essential for DNA integrity, is an important survival factor for cancer cells. We determined the crystal structure of the enzyme:a,b-imino-dUTP:Mg complex and performed equilibrium binding experiments in solution.Ordering of the C-terminus upon the active site induces close juxtaposition of the incoming nucleophile attacker water oxygen and the a-phosphorus of the substrate, decreasing their distance below the van der Waals limit. Complex interactions of the C-terminus with both substrate and product were observed via a specifically designed tryptophan sensor, suitable for further detailed kinetic and ligand binding studies. Results explain the key functional role of the C-terminus.

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Section: Resultssupporting

confidence: 65%

Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase

et al. 2007

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“…It was proven that DC p is an important indication of protein structural changes in response to different ligands since it correlates with the change in the surface area buried upon interaction (Jelesarov and Bosshard, 1999;Quesada-Soriano et al, 2007;Islam and Kumar, 2009). The change in heat capacity associated with ligand binding is believed to be associated with motion restriction and to changes in vibrational content on complex formation, and is proportional to the burial of the molecular surface, which generally correlates with a change in the solvent accessible surface area (Vamvaca et al, 2008).…”

Section: Contribution Of Structural Changes To the Thermodynamics Obsmentioning

confidence: 99%

Binding dynamics and energetic insight into the molecular forces driving nucleotide binding by guanylate kinase

Kandeel

Kitade

2010

J of Molecular Recognition

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Plasmodium deoxyguanylate pathways are an attractive area of investigation for future metabolic and drug discovery studies due to their unique substrate specificities. We investigated the energetic contribution to guanylate kinase substrate binding and the forces underlying ligand recognition. In the range from 20 to 35°C, the thermodynamic profiles displayed marked decrease in binding enthalpy, while the free energy of binding showed little changes. GMP produced a large binding heat capacity change of -356 cal mol(-1) K(-1), indicating considerable conformational changes upon ligand binding. Interestingly, the calculated ΔCp was -32 cal mol(-1) K(-1), indicating that the accessible surface area is not the central change in substrate binding, and that other entropic forces, including conformational changes, are more predominant. The thermodynamic signature for GMP is inconsistent with rigid-body association, while dGMP showed more or less rigid-body association. These binding profiles explain the poor catalytic efficiency and low affinity for dGMP compared with GMP. At low temperature, the ligands bind to the receptor site under the effect of hydrophobic forces. Interestingly, by increasing the temperature, the entropic forces gradually vanish and proceed to a nonfavorable contribution, and the interaction occurs mainly through bonding, electrostatic forces, and van der Waals interactions.

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“…As an example, we will use the binding of dUDP to trimeric dUTPase from Plasmodium falciparum (PfdUTPase) in glycerophosphate buffer at pH 7 and 25 ºC (Quesada-Soriano et al, 2007). Fig.…”

Section: Experimental Planningmentioning

confidence: 99%

Thermodynamics of Molecular Recognition by Calorimetry

Garcı́a-Fuentes¹,

Ramiro²,

Téllez-Sanz³

et al. 2011

Thermodynamics - Physical Chemistry of Aqueous Systems

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Plasmodium falciparum dUTPase: Studies on protein stability and binding of deoxyuridine derivatives

Cited by 11 publications

References 36 publications

Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase

Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase

Binding dynamics and energetic insight into the molecular forces driving nucleotide binding by guanylate kinase

Thermodynamics of Molecular Recognition by Calorimetry

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