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首页> 外文期刊>Journal of vector borne diseases >'DEAD-box' helicase from Plasmodium falciparum is active at wide pH and is schizont stage-specific.
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'DEAD-box' helicase from Plasmodium falciparum is active at wide pH and is schizont stage-specific.

机译：来自疟原虫的“死箱”螺旋酶在宽pH下活跃，是Schizont阶段特异性的。

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BACKGROUND & OBJECTIVES: DNA helicases catalyse unwinding of duplex DNA in an ATP-dependent manner and are involved in all the basic genetic processes. In order to study these important enzymes in the human malaria parasite we have recently cloned the first full-length 'DEAD-box' helicase gene from Plasmodium falciparum (3D7). In the present study, we report some of the important activities of the encoded protein. METHODS: We have expressed the P. falciparum helicase in Escherichia coli and characterised the encoded biochemically active helicase protein. The characterisation of the protein was carried out using radioactively labeled substrate and the standard strand displacement assay. The localisation of the enzyme was studied using immunofluorescence assay. RESULTS & CONCLUSION: P. falciparum helicase gene is 1551 bp in length and encodes for a protein consisting of 516 amino acid residues with a predicted molecular mass of 59.8 kDa. The protein is designated as Plasmodium falciparum DEAD-box helicase 60 kDa in size (PfDH60). Purified PfDH60 showed ATP and Mg2+ dependent DNA unwinding, ssDNA-dependent ATPase and ATP-binding activities. Interestingly, this is a unique helicase because it works at a wide pH range (from 5.0-10.0). The peak expression of PfDH60 is mainly in schizont stages of the development of P. falciparum, where DNA replication is active. The cell-cycle dependent expression suggests that PfDH60 may be involved in the process of DNA replication and distinct cellular processes in the parasite and this study should make an important contribution in our better understanding of DNA metabolic pathways in the parasite.
机译：背景和目标：DNA螺旋酶以ATP依赖性方式催化双链DNA的展开，并参与所有基本遗传过程。为了研究人疟疾寄生虫中的这些重要酶，我们最近克隆了来自疟原虫（3D7）的第一个全长的“死箱的螺旋酶基因”（3D7）。在本研究中，我们报告了编码蛋白的一些重要活动。方法：我们在大肠杆菌中表达了P.Malciparum Helicase，并表征了编码的生物化活性螺旋酶蛋白。使用放射性标记的基材和标准链位移测定来进行蛋白质的表征。使用免疫荧光测定研究酶的定位。结果与结论：P. falciparum Helicase基因的长度为1551bp，并编码由516个氨基酸残基组成的蛋白质，预测分子量为59.8kDa。该蛋白质被称为疟原虫死箱螺旋酶60kDa（PFDH60）。纯化的PFDH60显示ATP和MG2 +依赖性DNA斩波，SSDNA依赖性ATP酶和ATP结合活性。有趣的是，这是一个独特的螺旋酶，因为它在宽的pH范围内工作（从5.0-10.0）。 PFDH60的峰值表达主要是P. Falciparum的开发的Schizont阶段，其中DNA复制是活性的。细胞周期依赖性表达表明PFDH60可以参与寄生虫中DNA复制和不同细胞过程的过程，并且该研究应该在我们更好地了解寄生虫中的DNA代谢途径方面做出重要贡献。

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