A monomer form of the glutathione S-transferase Y7F mutant from Schistosoma japonicum at acidic pH

被引：6

作者：

Andújar-Sánchez, M ^{[1
]}

Clemente-Jimenez, JM ^{[1
]}

Rodriguez-Vico, F ^{[1
]}

Las Heras-Vazquez, FJ ^{[1
]}

Jara-Pérez, V ^{[1
]}

Cámara-Artigas, A ^{[1
]}

机构：

[1] Univ Almeria, Dept Quim Fis Bioquim & Quim Inorgan, Almeria 04120, Spain

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2004年 / 314卷 / 01期

关键词：

size exclusion chromatography; fluorescence; folding; isothermal titration calorimetry;

D O I：

10.1016/j.bbrc.2003.12.067

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：

Dissociation and unfolding of homodimeric glutathione S-transferase Y7F mutant from Schistosoma japonicum (SjGST-Y7F) were investigated at equilibrium using urea as denaturant. The conserved residue Tyr7 plays a central role in the catalytic mechanism and the mutation Tyr-Phe yields an inactive enzyme that is able to bind the substrate GSH with a higher binding constant than the wild type enzyme. Mutant SjGST-Y7F is a dimer at pH 6 or higher and a stable monomer at pH 5 that binds GSH (K value of 1.2 x 10(5) +/- 6.4 x 10(3) M-1 at pH 6.5 and 6.3 x 10(4) +/- 1.25 x 10(3) M-1 at pH 5). The stability of the SjGST-Y7F mutant was studied by urea induced unfolding techniques (DeltaG(W) = 13.86 +/- 0.63 kcal mol(-1) at pH 6.5 and DeltaG(W) = 11.22 +/- 0.25 kcal mol(-1) at pH 5) and the monomeric form characterized by means of size exclusion chromatography, fluorescence, and electrophoretic techniques. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：6 / 10

页数：5

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