Interrelationship between cationic and anionic forms of glutathione S-transferases of bovine ocular lens.
Since the eye is constantly exposed to potentially damaging chemical compounds present in the atmosphere and vascular system, we investigated the physiological role of glutathione S-transferase (GSH S-transferase) in detoxification mechanisms operative in the ocular lens. We have purified an anionic and a cationic GSH S-transferase from the bovine lens to homogeneity through a combination of gel filtration, ion-exchange and affinity chromatography. The anionic (pI 5.6) and cationic (pI 7.4) S-transferases were found to have distinct kinetic parameters (apparent Km and Vmax. pH optimum and energy of activation). However, both species were demonstrated to have similar molecular weights and amino acid compositions. Double-immunodiffusion and immunotitration studies showed that both lens S-transferases were immunologically similar. The very close similarity in amino acid compositions and immunological properties strongly indicates that these two transferases either originate from the same gene or at least share common antigenic determinants and originate from similar genes. The bovine lens GSH S-transferases had no glutathione peroxidase activity with either t-butyl hydroperoxide or cumene hydroperoxide as substrate. However, the antibody raised against the homogeneous anionic glutathione S-transferase from the bovine lens was found to precipitate both glutathione S-transferase and glutathione peroxidase activities out of solution in the supernatant of a crude bovine liver homogenate.
D000818:Animals; D000838:Anions; D002412:Cations; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D005979:Glutathione Peroxidase / Q000302:isolation & purification; D005982:Glutathione Transferase / Q000276:immunology / Q000302:isolation & purification*; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification*; D007700:Kinetics; D007908:Lens, Crystalline / Q000201:enzymology*; D013379:Substrate Specificity
D000838:Anions; D002412:Cations; D007527:Isoenzymes; D005979:Glutathione Peroxidase; D005982:Glutathione Transferase