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首页> 外文期刊>Biochemistry >The Ubiquitin-Proteasome Pathway Plays an Essential Role in Protelysis during Trypanosoma cruzi Remodeling
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The Ubiquitin-Proteasome Pathway Plays an Essential Role in Protelysis during Trypanosoma cruzi Remodeling

机译:泛素-蛋白酶体途径在克氏锥虫重塑过程中在蛋白水解中起重要作用

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Here, we document for the first time the presence of the 26S proteasome and the ubiquitin pathway in a protozoan parasite that is in an early branch in the eukaryotic lineage. The 26S proteasome of Trypanosoma cruzi epimastigotes was identified as a high molecular weight complex (1400 kDa) with an ATP-dependent chymotrypsin-like activity against the substrate Suc-LLVY-Amc. This activity was inhibited by proteasome inhibitors and showed same electrophorectic migration pattern as yeast 26S proteasome in nondenaturating gels. About 30 proteins in a range of 25 -110 kDa were detected in the purified T. cruzi 26S proteasome. Antibodies raised against the AAA family of A TPases from eukaryotic 26S proteasome and the T. cruzi 20S core specifically recognized components of T. cruzi 26S. To conflfffi the biological role of 26S in this primitive eukaryotic parasite, we analyzed the participation of the ubiquitin (Ub)-proteasome system in protein degradation during the time of parasite remodeling. Protein turnover in trypomastigotes was proteasome and A TP-dependent and was enhanced during the transformation of the parasites into amastigotes. If 20S proteasome activity is inhibited, ubiquitinated proteins accumulate in the parasites. As expected from the profound morphological changes that occur during transformation, cytos~eletal proteins associated with the fla~ellum are targets of the ubiquitin-proteasome pathway.
机译:在这里,我们首次记录了真核生物的早期分支中原生动物寄生虫中26S蛋白酶体和泛素途径的存在。克氏锥虫锥虫的26S蛋白酶体被鉴定为高分子量复合物(1400 kDa),具有针对底物Suc-LLVY-Amc的ATP依赖性胰凝乳蛋白酶样活性。该活性被蛋白酶体抑制剂抑制,并在非变性凝胶中显示出与酵母26S蛋白酶体相同的电泳迁移模式。在纯化的克氏锥虫26S蛋白酶体中检测到25种-110 kDa范围内的约30种蛋白质。来自真核生物26S蛋白酶体和克氏锥虫20S核心的针对A TPase AAA家族的抗体特异性识别了克氏锥虫26S的成分。为了确认26S在这种原始的真核生物寄生虫中的生物学作用,我们分析了在寄生虫重塑时泛素(Ub)-蛋白酶体系统参与蛋白质降解的过程。锥虫的蛋白质周转率是蛋白酶体和TP依赖性的,并且在寄生虫转化为amastigotes的过程中得到增强。如果抑制20S蛋白酶体活性,则泛素化的蛋白质会在寄生虫中积聚。正如从转化过程中发生的深刻形态变化所预期的那样,与黄素相关的细胞蛋白质是泛素-蛋白酶体途径的靶标。

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