title: Thermodynamic analysis of the binding of glutathione to glutathione S-transferase over a range of temperatures.
abstract: The binding properties of a glutathione S-transferase (EC 2.5.1.18) from Schistosoma japonicum to substrate glutathione (GSH) has been investigated by intrinsic fluorescence and isothermal titration calorimetry (ITC) at pH 6.5 over a temperature range of 15-30 degrees C. Calorimetric measurements in various buffer systems with different ionization heats suggest that protons are released during the binding of GSH at pH 6.5. We have also studied the effect of pH on the thermodynamics of GSH-GST interaction. The behaviour shown at different pHs indicates that at least three groups must participate in the exchange of protons. Fluorimetric and calorimetric measurements indicate that GSH binds to two sites in the dimer of 26-kDa glutathione S-transferase from Schistosoma japonicum (SjGST). On the other hand, noncooperativity for substrate binding to SjGST was detected over a temperature range of 15-30 degrees C. Among thermodynamic parameters, whereas DeltaG degrees remains practically invariant as a function of temperature, DeltaH and DeltaS degrees both decrease with an increase in temperature. While the binding is enthalpically favorable at all temperatures studied, at temperatures below 25 degrees C, DeltaG degrees is also favoured by entropic contributions. As the temperature increases, the entropic contributions progressively decrease, attaining a value of zero at 24.3 degrees C, and then becoming unfavorable. During this transition, the enthalpic contributions become progressively favorable, resulting in an enthalpy-entropy compensation. The temperature dependence of the enthalpy change yields the heat capacity change (DeltaCp degrees ) of -0.238 +/- 0.04 kcal per K per mol of GSH bound.
authors.affiliation: Dpto. Química Física, Bioquímica y Q. Inorgánica, Facultad de Ciencias Experimentales, Universidad de Almería, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Ortiz-Salmerón
authors.forename: Z
authors.initials: Z
authors.lastname: Yassin
authors.forename: M J
authors.initials: MJ
authors.lastname: Clemente-Jimenez
authors.forename: F J
authors.initials: FJ
authors.lastname: Las Heras-Vazquez
authors.forename: F
authors.initials: F
authors.lastname: Rodriguez-Vico
authors.forename: C
authors.initials: C
authors.lastname: Barón
authors.forename: L
authors.initials: L
authors.lastname: García-Fuentes
mesh_terms: D000818:Animals; D002151:Calorimetry; D005978:Glutathione / Q000737:chemistry* / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry* / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D015233:Models, Statistical; D011485:Protein Binding; D011522:Protons; D012549:Schistosoma japonicum / Q000737:chemistry / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics
chemical_list: D011522:Protons; D005982:Glutathione Transferase; D005978:Glutathione