Biblioteca de la Universidad Complutense de Madrid

Thermodynamic analysis of the binding of 5-fluoro-2'-deoxyuridine 5'-monophosphate to thymidylate synthase over a range of temperatures

Impacto



García Fuentes, Luis y Reche, Pedro A y López Mayorga, O y Santi, D V y González-Pacanowska, D. y Barón, C (1995) Thermodynamic analysis of the binding of 5-fluoro-2'-deoxyuridine 5'-monophosphate to thymidylate synthase over a range of temperatures. European Journal of Biochemistry / FEBS, 232 (2). pp. 641-5. ISSN 0014-2956

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Resumen

The binding of 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP) to Lactobacillus casei recombinant thymidylate synthase has been studied by isothermal titration microcalorimetry at pH 7.1 over the temperature range 16-35 degrees C. Calorimetric measurements in various buffer systems with different heats of ionization suggest that a proton uptake is involved in the binding process of the nucleotide. In the temperature range investigated, the mol protons bound/mol nucleotide increases as the temperature decreases. A model of two equal and independent sites fits well with the binding isotherms for thymidylate synthase. The binding constants, the changes in Gibbs energy, enthalpy, and entropy/site for FdUMP binding were calculated at each temperature. The results show that the binding is driven by both enthalpy and entropy contributions in the range 16-35 degrees C. The enthalpy changes become more negative as the temperature increases, with delta Cp = -170 +/- 20 J.K-1.(mol FdUMP bound)-1. The behavior of the system supports the observation that FdUMP binds to thymidylate synthase without producing profound conformational changes in the protein dimer.


Tipo de documento:Artículo
Palabras clave:Thymidylate synthase; 5-fluoro-2'-deoxyuridine 5'; Monophosphate; Microcalorimetry; Binding
Materias:Ciencias > Química > Biología molecular
Ciencias > Química > Bioquímica
Código ID:9354
Depositado:14 Aug 2009 10:56
Última Modificación:01 Sep 2010 10:57

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