title: Optimization of storage and stability of camel liver glutathione S-transferase.
abstract: Glutathione S-transferases (GSTs) are multifunctional enzymes and play an important role in cellular detoxification. Besides this, GSTs act as cytosolic carrier proteins that bind hydrophobic compounds such as heme, bilirubin, steroids, and polycyclic hydrocarbons. GST has great importance in biotechnology, as it is a target for vaccine and drug development and biosensors development for xenobiotics. Moreover, the GST tag has been extensively used for protein expression and purification. Until now, biophysical properties of camel liver GST have not been characterized. In the present study we have purified camel (Camelus dromedarius) liver GST to homogeneity in a single step by affinity chromatography with 23.4-fold purification and 60.6% yield. Our results showed that maximal activity of GST was at pH 6.5 and it was stable in the pH range of 5 to 10. The optimum temperature was 55°C and the Tm was 57°C. The chemical chaperone glycerol (3.3 M) was able to protect GST activity and aggregation against thermal denaturation by stabilizing the protein structure at 50 and 57°C, respectively. However, L-arginine (125 mM) did not protect GST against thermal stress. Far-ultraviolet circular dichroism (CD) spectra showed that glycerol protected the secondary structure of GST while L-arginine induced conformational changes under thermal stress. In conclusion, our studies on the GST stability suggest that glycerol works as a stabilizer and L-arginine acts as a destabilizer.
authors.affiliation: a Protein Research Chair, Department of Biochemistry, College of Science , King Saud University , Riyadh , Saudi Arabia.
authors.forename: Ajamaluddin
authors.initials: A
authors.lastname: Malik
authors.forename: Haseeb
authors.initials: H
authors.lastname: Jagirdar
authors.forename: Nayyar
authors.initials: N
authors.lastname: Rabbani
authors.forename: Mohd Shahnawaz
authors.initials: MS
authors.lastname: Khan
authors.forename: Anwar
authors.initials: A
authors.lastname: Ahmed
authors.forename: Abdulrahman M
authors.initials: AM
authors.lastname: Al-Senaidy
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Ismael
mesh_terms: D000818:Animals; D002162:Camelus; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D003600:Cytosol / Q000737:chemistry / Q000201:enzymology*; D005982:Glutathione Transferase / Q000737:chemistry* / Q000302:isolation & purification; D008099:Liver / Q000737:chemistry / Q000201:enzymology*
chemical_list: D005982:Glutathione Transferase
keywords: L-arginine; aggregation; chemical chaperone; circular dichroism; glutathione S-transferase; protein purification and characterization; protein stability; thermal unfolding