EFFECT OF IRON ON DISTRIBUTION OF SUPEROXIDE AND HYDROXYL RADICALS AS SEEN BY SPIN TRAPPING AND ON SUPEROXIDE-DISMUTASE ASSAY
EFFECT OF IRON ON DISTRIBUTION OF SUPEROXIDE AND HYDROXYL RADICALS AS SEEN BY SPIN TRAPPING AND ON SUPEROXIDE-DISMUTASE ASSAY
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DOI：
10.1111/j.1751-1097.1978.tb07001.x
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发表时间：
1978-01-01
影响因子：
3.3
通讯作者：
LEUTHAUSER, SWHC
中科院分区：
文献类型：
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作者：
BUETTNER, GR;OBERLEY, LW;LEUTHAUSER, SWHC
利用自旋陷阱5，5-二甲基-1-吡咯啉-1-氧化物，我们证明了在铁和/或铁-EDTA(乙二胺-四乙酸)存在下，黄嘌呤-黄嘌呤氧化酶体系中存在羟基。随着铁(或铁-EDTA)浓度的增加，O2-自旋加合物的强度减小，而OH的自旋加合物强度增加。然而，在反应混合物中使用二乙烯三胺五乙酸(DETAPAC)作为金属螯合剂，在保持O2-自旋加合物信号强度的同时，抑制了OH自旋加合物信号。使用EDTA消除金属离子的干扰效应，采用黄嘌呤氧化酶和氮蓝四唑测定超氧化物歧化酶，引入了铁存在的伪影。使用DETAPAC作为金属螯合剂，可以消除包括铁在内的金属离子对测定的干扰。因此，即使在样品中存在铁含量的变化时，也可以对测量的超氧化物歧化酶活性进行比较。
Using the spin trap 5,5‐dimethyl‐1‐pyrroline‐1‐oxide we have demonstrated the presence of OH in the xanthine‐xanthine oxidase system when iron and/or iron‐EDTA (ethylenediamine‐tetraacetic acid) is present. With increasing iron (or iron‐EDTA) concentration the intensity of the O2‐spin adduct decreased while that of OH increased. However, use of diethylenetriaminepentaacetic acid (DETAPAC) as a metal chelator in the reaction mixture suppressed the OH spin adduct signal while maintaining the intensity of the signal from the O2‐spin adduct.Use of EDTA to eliminate the interfering effects of metal ions in the superoxide dismutase assay employing xanthine oxidase and nitroblue tetrazolium introduces an artifact from the iron present. The interference in the assay from metal ions, including iron, can be eliminated with use of DETAPAC as a metal chelator. Thus, it is possible to make comparisons of measured superoxide dismutase activities even when there are variations in the amount of iron present in the samples.