Structural and Functional Characterization of Camelus dromedarius Glutathione Transferase M1-1

Glutathione transferases (GSTs; EC. 2.5.1.18) are a large family of multifunctional enzymes that play crucial roles in the metabolism and inactivation of a broad range of xenobiotic compounds. In the present work, we report the kinetic and structural characterization of the isoenzyme GSTM1-1 from Camelus dromedarius (CdGSTM1-1). The CdGS tau M1-1 was expressed in E. coli BL21 (DE3) and was purified by affinity chromatography. Kinetics analysis showed that the enzyme displays a relative narrow substrate specificity and restricted ability to bind xenobiotic compounds. The crystal structures of CdGS tau M1-1 were determined by X-ray crystallography in complex with the substrate (GSH) or the reaction product (S-p-nitrobenzyl-GSH), providing snapshots of the induced-fit catalytic mechanism. The thermodynamic stability of CdGSTM1-1 was investigated using differential scanning fluorimetry (DSF) in the absence and in presence of GSH and S-p-nitrobenzyl-GSH and revealed that the enzyme's structure is significantly stabilized by its ligands. The results of the present study advance the understanding of camelid GST detoxification mechanisms and their contribution to abiotic stress adaptation in harsh desert conditions.

Automated summary

This study reported the kinetic and structural characterization of GSTM1-1 from Camelus dromedarius, revealing its narrow substrate specificity and limited ability to bind xenobiotic compounds. The crystal structures of CdGS tau M1-1 in complex with the substrate or the reaction product provided insights into the induced-fit catalytic mechanism. Furthermore, the thermodynamic stability of CdGSTM1-1 was investigated, showing its significant stabilization by ligands. These findings advance our understanding of the detoxification mechanisms and stress adaptation of camelid GSTs.