title: Preliminary studies on the renaturation of denatured catfish (Clarias gariepinus) glutathione transferase.
abstract: Purified juvenile catfish (Clarias gariepinus) glutathione transferase (cgGST) was denatured in vitro and renatured in the absence and presence of different concentrations of endogenous or xenobiotic model substrates. Protein transitions during unfolding and refolding were monitored by activity measurement as well as changes in protein conformation using UV difference spectra at 230 nm. Gdn-HCl at 0.22 M caused 50 % inactivation of the enzyme and at 1.1 M, the enzyme was completely unfolded. Refolding of cgGST main isozyme was not completely reversible at higher concentrations of Gdn-HCl and is dependent on protein concentration. An enzyme concentration of 30 μg/ml yielded 40 % percentage residual activity in the presence of glutathione (GSH), regardless of the concentration that was present as opposed to 30 % obtained in its absence. The xenobiotic model substrate, lindane, appears to have no effect on the refolding of the enzyme. In summary, our results show that GSH assists in the refolding of cgGST in a concentration-independent manner and may be involved in the same function in vivo whereas the xenobiotic model substrate does not.
authors.affiliation: Department of Biochemistry, Obafemi Awolowo University, Ile-Ife, Nigeria.
authors.forename: Yetunde Adedolapo
authors.initials: YA
authors.lastname: Ojopagogo
authors.forename: Isaac Olusanjo
authors.initials: IO
authors.lastname: Adewale
authors.forename: Adeyinka
authors.initials: A
authors.lastname: Afolayan
mesh_terms: D000818:Animals; D002397:Catfishes / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism*; D019791:Guanidine; D001556:Hexachlorocyclohexane / Q000378:metabolism / Q000633:toxicity*; D011489:Protein Denaturation; D020673:Protein Renaturation; D015262:Xenobiotics / Q000378:metabolism / Q000633:toxicity*
chemical_list: D015262:Xenobiotics; D001556:Hexachlorocyclohexane; D005982:Glutathione Transferase; D005978:Glutathione; D019791:Guanidine