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KMID : 1011420150200010075
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Journal of Korean Ophthalmic Optics Society
2015 Volume.20 No. 1 p.75 ~ p.81
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The Relationship between Structural Denaturation of Antioxidative Enzymes and Their Enzyme Activity due to Repeated Exposure to UV-A
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박미정:Park Mi-Jung
유효정:Yoo Hyo-Jung/김종찬:Kim Jong-Chan/김소라:Kim So-Ra
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Abstract
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Purpose: The present study was conducted to investigate whether the changes in structure and activity of antioxidative enzymes, superoxide dismutase(SOD) and catalase(CAT) present in the eyes appeared when they were repeatedly exposed to UV-A, and reveal the correlation of these changes.
Methods: Each enzyme solution was prepared from the standardized SOD and CAT, and repeatedly exposed to UV-A of 365 min under the condition of 30 minutes, 1 hour and 2 hours a day over 1, 2, 3, 4 and 5 days. Structural denaturation of SOD and CAT induced by repeat UV-A irradiation was confirmed by the electrophoretic analysis, and their enzyme activity
was determined by the colorimetric assay using the proper assay kit.
Results: SOD exposed repeatedly to UV-A showed the polymerization pattern through the electrophoretic analysis when it was repeatedly exposed under the condition of at least 1 hour a day however, the change of its activity was found to be less than 12%. On the other hand, CAT repeatedly exposed to UV-A showed reduced size of the electrophoretic band which indicated a structure denaturation and its activity was significantly decreased. In the case of that the repeat exposure time was longer, CAT activity was completely lost even though some enzyme band was shown in the electrphoretic analysis.
Conclusions: From these results, it was revealed that the degree and pattern in structural denaturation of antioxidative enzymes differently appeared according to the type of enzyme, and the degree of structural denaturation was not always consistent with the reduction in enzyme activity.
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KEYWORD
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UV irradiation, Antioxidative enzyme, Superoxide dismutase, Catalase, Structural denaturation, Enzyme activity
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