Stability of the anti-oxidative enzymes in aqueous and detergent solution.
Activities of the anti-oxidative enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase were studied in rat tissues to determine the ability of detergents both to solubilize the enzymes and also to stabilize enzyme activity. Rat brain, heart and liver were homogenized in 0.1M KCl, 0.1% sodium dodecyl sulfate, 0.1% lubrol, or 0.1% cetyl-trimethylammonium bromide. In general lubrol was more effective than the other solutions in solubilizing GPx and catalase. Lubrol and 0.1M KCl were equally effective in solubilizing SOD. The highest enzyme activities were (1) SOD: 2484 ng/mg (brain), 2501 ng/mg (heart), and 5586 ng/mg (liver); (2) GPx: 224 mU/mg (brain), 1870 mU/mg (heart), and 7332 mU/mg (liver); (3) catalase: 2.8 mU/mg (brain), 10.6 mU/mg (heart), and 309 mU/mg (liver). While cetyl trimethylammonium bromide is marginally better than sodium dodecyl sulfate in solubilizing active enzyme, neither ionic detergent has any advantage over lubrol or 0.1M KCl. For catalase and GPx, enzyme activity loss with time is biphasic. After initial, rapid activity loss (1-5 days for GPx and 7-10 days for catalase) the differences noted among the homogenizing solutions disappear and very little if any activity loss is noted over the next 2-3 weeks. For catalase and GPx, only baseline enzyme activity from t = 0-3 weeks is found in the most chaotropic solution, 0.1% sodium dodecyl sulfate while biphasic activity loss is most pronounced in 0.1% lubrol. These results may indicate active GPx and catalase species stabilized by a lipid-like environment. Correlating in vitro catalase or GPx measurements with in vivo anti-oxidative protection may underestimate tissue defences.
Mailer
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Faculty of Science, Athabasca University, Alberta, Ontario, Canada.
Del Maestro
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RF
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001921:Brain / Q000201:enzymology; D002374:Catalase / Q000187:drug effects / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D005979:Glutathione Peroxidase / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D011489:Protein Denaturation / Q000187:drug effects; D051381:Rats; D012996:Solutions; D013482:Superoxide Dismutase / Q000187:drug effects / Q000378:metabolism*; D014867:Water
D000975:Antioxidants; D003902:Detergents; D012996:Solutions; D014867:Water; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase