Crystallographic and thermodynamic analysis of the binding of S-octylglutathione to the Tyr 7 to phe mutant of glutathione S-transferase from Schistosoma japonicum

被引:22
作者
Andújar-Sánchez, M
Smith, AW
Clemente-Jimenez, JM
Rodriguez-Vico, F
Heras-Vazquez, FJL
Jara-Pérez, V
Cámara-Artigas, A
机构
[1] Univ Almeria, Dept Quim Fis Bioquim & Quim Inorgan, Almeria, Spain
[2] Arizona State Univ, Dept Chem & Biochem, Tempe, AZ 85287 USA
关键词
D O I
10.1021/bi0483110
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glutathione S-transferases are a family of multifunctional enzymes involved in the metabolism of drugs and xenobiotics. Two tyrosine residues, Tyr 7 and Tyr 111, in the active site of the enzyme play an important role in the binding and catalysis of substrate ligands. The crystal structures of Schistosoma japonicum glutathione S-transferase tyrosine 7 to phenylalanine mutant [SjGST(Y7F)] in complex with the substrate glutathione (GSH) and the competitive inhibitor S-octylglutathione (S-octyl-GSH) have been obtained. These new structural data combined with fluorescence spectroscopy and thermodynamic data, obtained by means of isothermal titration calorimetry, allow for detailed characterization of the ligand-binding process. The binding of S-octyl-GSH to SjGST(Y7F) is enthalpically and entropically driven at temperatures below 30degreesC. The stoichiometry of the binding is one molecule of S-octyl-GSH per mutant dimer, whereas shorter alkyl derivatives bind with a stoichiometry of two molecules per mutant dimer. The SjGST(Y7F)(.)GSH structure showed no major structural differences compared to the wild-type enzyme. In contrast, the structure of SjGST(Y7F)S-.-octyl-GSH showed asymmetric binding of S-octyl-GSH. This lack of symmetry is reflected in the lower symmetry space group of the SjGST(Y7F)S-.-octyl-GSH crystals (P6(3)) compared to that of the SjGST(Y7F)(.)GSH crystals (P6(3)22). Moreover, the binding of S-octyl-GSH to the A subunit is accompanied by conformational changes that may be responsible for the lack of binding to the B subunit.
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页码:1174 / 1183
页数:10
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