Abstract
Glutathione transferase (GST, EC 2.5.1.18) is a primary line of defense against toxicities of electrophile compounds and oxidative stress and therefore is involved in stress-response and cell detoxification. In the present study, we investigated the catalytic and structural properties of the glutathione transferase (GST) isoenzyme P1-1 from Camelus dromedarius (CdGSTP1-1). Recombinant CdGSTP1-1 was produced in Escherichia coli BL21(DE3) and purified to electrophoretic homogeneity. Kinetic analysis revealed that CdGSTP1-1 displays broad substrate specificity and shows high activity towards halogenated aryl-compounds, isothiocyanates and hydroperoxides. Computation analysis and structural comparison of the catalytic and ligand binding sites of CdGSTP1-1 with other pi class GSTs allowed the identification of major structural variations that affect the active site pocket and the catalytic mechanism., Affinity labeling and kinetic inhibition studies identified key regions that form the ligandin-binding site (L-site) and gave further insights into the mechanism of non-substrate ligand recognition. The results of the present study provide new information into camelid detoxifying mechanism and new knowledge into the diversity and complex enzymatic functions of GST superfamily.