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首页> 外文期刊>Parasitology >Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae)
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Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae)

机译：采血单生大肠棘突E（Eudiplozoon nipponicum）（Heteronchoinea：Diplozoidae）的主要酸性内肽酶

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摘要

In parasitic flatworms, acid endopeptidases are involved in crucial processes, including digestion, invasion, interactions with the host immune system, etc. In haematophagous monogeneans, however, no solid information has been available about the occurrence of these enzymes. Here we aimed to identify major cysteine and aspartic endopeptidase activities in Eudiplozoon nipponicum, an invasive haematophagous parasite of common carp. Employing biochemical, proteomic and molecular tools, we found that cysteine peptidase activities prevailed in soluble protein extracts and excretory/secretory products (ESP) of E. nipponicum; the major part was cathepsin L-like in nature supplemented with cathepsin B-like activity. Significant activity of the aspartic cathepsin D also occurred in soluble protein extracts. The degradation of haemoglobin in the presence of ESP and worm protein extracts was completely inhibited by a combination of cysteine and aspartic peptidase inhibitors, and diminished by particular cathepsin L, B and D inhibitors. Mass spectrometry revealed several tryptic peptides in ESP matching to two translated sequences of cathepsin L genes, which were amplified from cDNA of E. nipponicum and bioinformatically annotated. The dominance of cysteine peptidases of cathepsin L type in E. nipponicum resembles the situation in, e.g. fasciolid trematodes.
机译：在寄生性扁虫中，酸性内肽酶参与关键的过程，包括消化，侵袭，与宿主免疫系统的相互作用等。然而，在单食性嗜血菌中，尚无关于这些酶发生的可靠信息。在这里，我们旨在确定Eudiplozoon nipponicum（一种鲤鱼的侵入性食血性寄生虫）中的主要半胱氨酸和天冬氨酸内肽酶活性。利用生化，蛋白质组学和分子学工具，我们发现半胱氨酸肽酶活性普遍存在于大肠埃希菌的可溶性蛋白提取物和排泄/分泌产物（ESP）中。自然界中主要是组织蛋白酶L样，并补充了组织蛋白酶B样活性。天冬氨酸组织蛋白酶D的显着活性也发生在可溶性蛋白提取物中。在半胱氨酸和天冬氨酸肽酶抑制剂的组合下，在ESP和蠕虫蛋白提取物存在下，血红蛋白的降解被完全抑制，而特定的组织蛋白酶L，B和D抑制剂则抑制了这种降解。质谱分析显示，ESP中有多个胰蛋白酶肽与组织蛋白酶L基因的两个翻译序列相匹配，这两个序列是从N. nipponicum的cDNA中扩增得到的，并进行了生物信息学注释。 Nipponicum中组织蛋白酶L型半胱氨酸肽酶的优势类似于例如筋膜吸虫。

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