A general strategy to solve the phase problem in RNA crystallography
A general strategy to solve the phase problem in RNA crystallography
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DOI：
10.1016/j.str.2007.06.003
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发表时间：
2007-07-01
期刊：
影响因子：
5.7
通讯作者：
Kieft, Jeffrey S.
中科院分区：
文献类型：
--
作者：
Keel, Amanda Y.;Rambo, Robert P.;Kieft, Jeffrey S.
生物学上重要的RNA分子的X射线晶体学一直受到技术挑战的阻碍，包括寻找重原子衍生物以获得高质量的实验相位信息。现有的技术有缺点，限制了解决重要新结构的速度。为了解决这个问题，我们开发了一种可靠的方法，可以将重原子特异性地定位到几乎任何RNA上。通过解决G中心点U摆动对阳离子结合基序的十三种变体的晶体结构，我们已经确定了一个版本，当插入到RNA螺旋中时，引入了适合定相的高占用率阳离子结合位点。这种“定向浸泡”策略可以完全整合到现有的RNA晶体学方法中，从而可能提高重要结构的求解速率，并促进使用Cu-K α辐射进行结构的常规求解。这种方法已经被用来解决几个晶体结构。
X-ray crystallography of biologically important RNA molecules has been hampered by technical challenges, including finding heavy-atom derivatives to obtain high-quality experimental phase information. Existing techniques have drawbacks, limiting the rate at which important new structures are solved. To address this, we have developed a reliable means to localize heavy atoms specifically to virtually any RNA. By solving the crystal structures of thirteen variants of the G center dot U wobble pair cation binding motif, we have identified a version that when inserted into an RNA helix introduces a high-occupancy cation binding site suitable for phasing. This "directed soaking" strategy can be integrated fully into existing RNA crystallography methods, potentially increasing the rate at which important structures are solved and facilitating routine solving of structures using Cu-K alpha radiation. This method already has been used to solve several crystal structures.