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反刍动物胃肠道线虫中蛋白水解酶(蛋白酶)的存在与释放研究。

Studies on the presence and release of proteolytic enzymes (proteinases) in gastro-intestinal nematodes of ruminants.

作者信息

Knox D P, Jones D G

机构信息

Moredun Research Institute, Edinburgh, U.K.

出版信息

Int J Parasitol. 1990 Apr;20(2):243-9. doi: 10.1016/0020-7519(90)90106-w.

Abstract

The proteolytic activity of worm homogenates prepared from the third stage larval (L3) and adult stages of the ovine gastro-intestinal nematodes, Nematodirus battus, Ostertagia circumcincta, Trichostrongylus colubriformis, Trichostrongylus vitrinus and Haemonchus contortus, and the rodent intestinal nematode, Nippostrongylus brasiliensis, has been examined using the protein substrates azocasein, azocoll and elastin-orcein. Activity detected in third stage larvae was usually higher than that observed in the adult. Species and stage differences were demonstrated. The in vitro release of proteolytic activity, detected using protein substrates and specific low molecular weight peptide substrates, was, similarly, shown to exhibit a degree of species and stage specificity. Acetylcholinesterase and lactate dehydrogenase activities were determined as reference 'secretory' and 'non-secretory' enzymes, respectively. Three separate peaks of 'tryptic' activity were detected following anion-exchange chromatography of culture fluids derived from adult Ostertagia circumcincta. These peaks could be ascribed to different proteinase classes on the basis of inhibitor sensitivity.

摘要

利用蛋白质底物偶氮酪蛋白、偶氮胶体和弹性蛋白orcein,检测了从绵羊胃肠道线虫巴贝斯细颈线虫、环形奥斯特线虫、蛇形毛圆线虫、玻璃样毛圆线虫和捻转血矛线虫的第三期幼虫(L3)和成虫阶段,以及啮齿动物肠道线虫巴西日圆线虫制备的虫体匀浆的蛋白水解活性。在第三期幼虫中检测到的活性通常高于成虫中观察到的活性。证明了物种和阶段差异。同样,使用蛋白质底物和特定的低分子量肽底物检测到的蛋白水解活性的体外释放显示出一定程度的物种和阶段特异性。分别测定乙酰胆碱酯酶和乳酸脱氢酶活性作为参考“分泌性”和“非分泌性”酶。对成年环形奥斯特线虫培养液进行阴离子交换色谱分析后,检测到三个独立的“胰蛋白酶样”活性峰。根据抑制剂敏感性,这些峰可归因于不同的蛋白酶类别。

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