A NMR guided approach for CsrA-RNA crystallization
A NMR guided approach for CsrA-RNA crystallization
复制标题
DOI：
10.1007/s10858-013-9712-3
复制
发表时间：
2013-05-01
影响因子：
2.7
通讯作者：
Gronenborn, Angela M.
中科院分区：
文献类型：
--
作者：
Koharudin, Leonardus M. I.;Boelens, Rolf;Gronenborn, Angela M.
蛋白质-核酸复合物的结构测定仍然是一项具有挑战性的任务。在这里，我们提出了一种简单的方法来产生晶体的csra -核酸复合物，完全由核磁共振波谱（NMR）的结果指导。使用缺乏13个非必需c端残基的结构，可以证明与DNA的有效结合。一个CsrA二聚体与两个DNA寡核苷酸相互作用，类似于之前与RNA的发现。此外，对CsrA-DNA复合物的核磁共振研究是成功定位条件的基础，适合获得CsrA-DNA复合物的晶体。我们的结果可能对那些RNA在蛋白质-核酸复合物中可能被DNA取代的情况有用。
Structure determination of protein-nucleic acid complexes remains a challenging task. Here we present a simple method for generating crystals of a CsrA-nucleic acid complex, guided entirely by results from nuclear magnetic resonances spectroscopy (NMR) spectroscopy. Using a construct that lacks thirteen non-essential C-terminal residues, efficient binding to DNA could be demonstrated. One CsrA dimer interacts with two DNA oligonucleotides, similar to previous findings with RNA. Furthermore, the NMR study of the CsrA-DNA complex was the basis for successfully homing in on conditions that were suitable for obtaining crystals of the CsrA-DNA complex. Our results may be useful for those cases where RNA in protein-nucleic acid complexes may be replaced by DNA.