Hybrid Methods in Iron-Sulfur Cluster Biogenesis.
Hybrid Methods in Iron-Sulfur Cluster Biogenesis.
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DOI：
10.3389/fmolb.2017.00012
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发表时间：
2017
影响因子：
5
通讯作者：
Pastore A
中科院分区：
文献类型：
--
作者：
Prischi F;Pastore A
混合方法结合并整合了几种生物化学和生物物理技术，在过去的二十年中迅速发展起来，提供了一种方法来获得更全面的蛋白质和分子复合物的大小和复杂性，否则是不容易负担得起的。在这里，我们回顾了一种基于NMR， SAXS，位点定向诱变和分子对接的混合方法的使用，我们已经开发出用于确定弱相互作用分子复合物结构的方法。我们应用这项技术来深入了解分子机器中蛋白质之间形成的复合物的结构，这些复合物产生了必需的铁硫簇假基。我们的结果得到了x射线结构和其他采用相同方法的小组的验证。我们讨论了我们的方法的优点和局限性，并提出了新的途径，可以改进它。
Hybrid methods, which combine and integrate several biochemical and biophysical techniques, have rapidly caught up in the last twenty years to provide a way to obtain a fuller description of proteins and molecular complexes with sizes and complexity otherwise not easily affordable. Here, we review the use of a robust hybrid methodology based on a mixture of NMR, SAXS, site directed mutagenesis and molecular docking which we have developed to determine the structure of weakly interacting molecular complexes. We applied this technique to gain insights into the structure of complexes formed amongst proteins involved in the molecular machine, which produces the essential iron-sulfur cluster prosthetic groups. Our results were validated both by X-ray structures and by other groups who adopted the same approach. We discuss the advantages and the limitations of our methodology and propose new avenues, which could improve it.