Resolution and characterization of the structural polymorphism of a single quadruplex-forming sequence.
Resolution and characterization of the structural polymorphism of a single quadruplex-forming sequence.
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DOI：
10.1093/nar/gkq166
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发表时间：
2010-08
影响因子：
14.9
通讯作者：
Trent JO
中科院分区：
文献类型：
--
作者：
Dailey MM;Miller MC;Bates PJ;Lane AN;Trent JO
四链形成序列的显著结构多态一直是阐明四链折叠的一个相当大的障碍。序列修改通常被用来干扰并据称从用于核磁共振(NMR)或X射线结晶学分析的折叠集合中选择特定形式。在这里，我们报告了一种简单的层析技术，它可以分离单个折叠，而不需要修改序列。根据各种常见的生物物理技术，序列d(GGTGGTGGGGTGTGTGGTGGGGGGGGGGGGGTGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG然而，核磁共振和层析显示，这种寡核苷酸至少产生8个单体四链物种，在室温下相互转换非常缓慢。我们使用光谱、流体力学和热力学技术相结合的方法来评估混合物和单个物种的物理化学性质。这些物种具有几乎相同的热力学、流体力学和电泳学性质，但核磁共振和圆二色谱(CD)光谱以及动力学稳定性有显著不同。这些结果表明，简单的标准低分辨率技术并不总是用于四链折叠测定或质量控制目的，简单的热力学分析不能直接提供可解释的热力学参数。
The remarkable structural polymorphism of quadruplex-forming sequences has been a considerable impediment in the elucidation of quadruplex folds. Sequence modifications have commonly been used to perturb and purportedly select a particular form out of the ensemble of folds for nuclear magnetic resonance (NMR) or X-ray crystallographic analysis. Here we report a simple chromatographic technique that separates the individual folds without need for sequence modification. The sequence d(GGTGGTGGTGGTTGTGGTGGTGGTGG) forms a compact quadruplex according to a variety of common biophysical techniques. However, NMR and chromatography showed that this oligonucleotide produces at least eight monomeric quadruplex species that interconvert very slowly at room temperature. We have used a combination of spectroscopic, hydrodynamic and thermodynamic techniques to evaluate the physicochemical properties of the mixture and the individual species. These species have almost identical thermodynamic, hydrodynamic and electrophoretic properties, but significantly different NMR and circular dichroism (CD) spectra, as well as kinetic stability. These results demonstrate that simple standard low-resolution techniques cannot always be used for quadruplex fold determination or quality control purposes, and that simple thermodynamic analysis does not directly provide interpretable thermodynamic parameters.
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影响因子：
5.7
作者：
Girvan, Allicia C.;Teng, Yun;Bates, Paula J.
通讯作者：
Bates, Paula J.
影响因子：
14.9
作者：
Lane AN;Chaires JB;Gray RD;Trent JO
通讯作者：
Trent JO
DOI：
10.1111/j.1742-4658.2009.07462.x
发表时间：
2010-03
期刊：
The FEBS journal
影响因子：
--
作者：
Chaires JB
通讯作者：
Chaires JB
影响因子：
15
作者：
Green, Jeremy J.;Ladame, Sylvain;Ying, Liming;Klenerman, David;Balasubramanian, Shankar
通讯作者：
Balasubramanian, Shankar
影响因子：
2.9
作者：
Hud, NV;Smith, FW;Feigon, J
通讯作者：
Feigon, J