Cu/Zn superoxide dismutase in excretory-secretory products of the human hookworm Necator americanus -: An electron paramagnetic spectrometry study

被引:21
作者
Taiwo, FA [1 ]
Brophy, PM
Pritchard, DI
Brown, A
Wardlaw, A
Patterson, LH
机构
[1] De Montfort Univ, Sch Pharm & Pharmaceut Sci, Leicester LE1 9BH, Leics, England
[2] Univ Leicester, Sch Med, Glenfield Hosp, Leicester, Leics, England
[3] Univ Nottingham, Dept Life Sci, Nottingham NG7 2RD, England
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1999年 / 264卷 / 02期
关键词
electron spin resonance spectrometry; EPR spectrometry; helminths; superoxide dismutase;
D O I
10.1046/j.1432-1327.1999.00626.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
EPR spectrometry was used to investigate the effect of excretory/secretory product from Necator americanus on superoxide radical anions generated by xanthine/xanthine oxidase as a measure of excretory/secretory product superoxide dismutase activity. Using 1,1',5,5'-dimethylpyrollidine-N-oxide (DMPO) as a superoxide spin-trapping agent a 12-line EPR spectrum characteristic of the DMPO-OOH adduct was observed to decrease in the presence of excretory/secretory product. Superoxide dismutase activity was proportional to excretory/secretory protein concentration, was inhibited with cyanide treatment and was progressively destroyed with increasing time of heat denaturation of excretory/secretory product. Using a purpose-built chamber the superoxide dismutase activity of excretory/secretory product from live worms in culture was shown to accumulate with time to a maximum at 4 h. The electron paramagnetic resonance spectrum obtained for the frozen excretory/secretory product of N. americanus recorded at 77 K is typical of Cu(II) in a protein matrix. The results are consistent with the presence of an active Cu/Zn superoxide dismutase in excretory/secretory product from N. americanus and demonstrate a method for the unequivocal determination of the fate of superoxide anions in the presence of live worms.
引用
收藏
页码:434 / 438
页数:5
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