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Intrinsic Disorder-Based Design of Stabilizing Disulphide Bridge in G alpha o Protein

机译:基于内在疾病的稳定G alpha o蛋白中二硫键的设计

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In this study, we have used an approach that allows us to determine in what region of the polypeptide chain of protein it is required to insert a disulphide bond in order to stabilize it. In our previous paper [Melnik et al., JBSD. 2012] it was proposed that to search for a "weak" site in the protein, it is possible to use programs (for example, PONDR-FIT and IsUnstruct) finding intrinsic disorder protein regions. We suggested that in structured globular proteins, such programs predict not protein regions in the polypeptide chain disordered under native conditions, but "weakened", feebly stabilized ones. Accordingly, an artificial introduction of SS-bridges using mutations in such regions would reliably result in the protein stabilization. We have taken advantage of this approach to stabilize protein G alpha o from Drosophila melanogaster. The designed SS-bridge increased by 4 degrees the melting temperature of one domain of protein G alpha o.
机译:在这项研究中,我们使用了一种方法,该方法可以确定在多肽多肽链的哪个区域插入二硫键以使其稳定。在我们以前的论文中[Melnik等,JBSD。 2012]提出要在蛋白质中搜索“弱”位点,可以使用程序(例如PONDR-FIT和IsUnstruct)查找内在的失调蛋白质区域。我们建议,在结构化的球形蛋白质中,此类程序预测的不是天然条件下无序的多肽链中的蛋白质区域,而是“弱化的”,微弱稳定的蛋白质区域。因此,在这种区域中使用突变人工引入SS桥将可靠地导致蛋白质稳定。我们已经利用这种方法来稳定果蝇中的G蛋白。设计的SS桥将蛋白G alpha o的一个结构域的解链温度提高了4度。

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