A calorimetric study of the binding of S-alkylglutathiones to glutathione S-transferase.
The binding of three competitive glutathione analogue inhibitors (S-alkylglutathione derivatives) to glutathione S-transferase from Schistosoma japonicum, SjGST, has been investigated by isothermal titration microcalorimetry 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 no protons are exchanged during the binding of S-alkylglutathione derivatives. Thus, at pH 6.5, the protons released during the binding of substrate may be from its thiol group. Calorimetric analyses show that S-methyl-, S-butyl-, and S-octylglutathione bind to two equal and independent sites in the dimer of SjGST. The affinity of these inhibitors to SjGST is greater as the number of methylene groups in the hydrocarbon side chain increases. In all cases studied, Delta G(0) remains invariant as a function of temperature, while Delta H(b) and Delta S(0) both decrease as the temperature increases. The binding of three S-alkylglutathione derivatives to the enzyme is enthalpically favourable at all temperatures studied. The temperature dependence of the enthalpy change yields negative heat capacity changes, which become less negative as the length of the side chain increases.
Ortiz-Salmerón
E
E
Departamento Química Física, Bioquímica y Q. Inorgánica, Facultad de Ciencias Experimentales, Universidad de Almería, La Cañada de San Urbano, 04120 Almería, Spain.
Yassin
Z
Z
Clemente-Jimenez
M J
MJ
Las Heras-Vazquez
F J
FJ
Rodriguez-Vico
F
F
Barón
C
C
García-Fuentes
L
L
D002021:Buffers; D002151:Calorimetry; D003461:Crystallography; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000494:pharmacology; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011485:Protein Binding; D011522:Protons; D013696:Temperature; D013816:Thermodynamics; D017186:Titrimetry
D002021:Buffers; D004791:Enzyme Inhibitors; D011522:Protons; C059196:S-octylglutathione; D005982:Glutathione Transferase; D005978:Glutathione