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首页> 外文期刊>The Korean Journal of Parasitology >Cytotoxicity of a cysteine proteinase of adult Clonorchis sinensis
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Cytotoxicity of a cysteine proteinase of adult Clonorchis sinensis

机译：成年华支睾吸虫半胱氨酸蛋白酶的细胞毒性

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摘要

To clarify the correlation of the proteinase activity with pathogenicity of Clonorchis sinensis, the proteinase activity either in excretory-secretory products (ESP) or in crude extracts of adult C. sinensis was examined. Substrate gel electrophoresis of the ESP and crude extracts revealed four distinct enzyme bands, which were differently inhibited by the specific proteinase inhibitors. The proteinase of the ESP with molecular mass of 24 kDa, was purified 23- fold with 14.5% yield by spectra gel ACA 44 gel filtration. It exhibited optimal pH at 7.5 in sodium phosphate (0.1 M). Its activity was inhibited specifically by N-ethylmaleimide (NEM) and antipain whereas potentiated 1.9 folds in the presence of 5 mM dithiothreitol (DTT). Cytotoxicity of the proteinase increased in a dose-dependent manner up to 120 μg/ml while reduced by NEM and antipain, indicating that cysteine proteinase was responsible for the cytotoxicity. This result shows that the 24 kDa cysteine proteinase is deeply correlated with the pathogenicity of C. sinensis infection.
机译：为了阐明蛋白酶活性与中华支睾吸虫病的致病性之间的关系，研究了成虫分泌物（ESP）或成年梭菌粗提物中的蛋白酶活性。 ESP和粗提物的底物凝胶电泳显示出四个不同的酶带，这些酶带被特定的蛋白酶抑制剂不同地抑制。通过光谱凝胶ACA 44凝胶过滤，将分子量为24 kDa的ESP蛋白酶纯化23倍，收率14.5％。在磷酸钠（0.1 M）中，其在7.5的最佳pH值。它的活性被N-乙基马来酰亚胺（NEM）和抗痛剂特异性抑制，而在5 mM二硫苏糖醇（DTT）存在下增强了1.9倍。蛋白酶的细胞毒性以剂量依赖性方式增加，直至120μg/ ml，而被NEM和抗痛药降低，表明半胱氨酸蛋白酶是造成细胞毒性的原因。该结果表明24kDa半胱氨酸蛋白酶与中华假丝酵母感染的致病性密切相关。

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