Analysis of oligonucleotide DNA binding and sedimentation properties of montmorillonite clay using ultraviolet light spectroscopy.
Analysis of oligonucleotide DNA binding and sedimentation properties of montmorillonite clay using ultraviolet light spectroscopy.
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DOI：
10.1021/bm800970v
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发表时间：
2009-01-12
影响因子：
6.2
通讯作者：
Lewis, L. Kevin
中科院分区：
文献类型：
--
作者：
Beall, Gary W.;Sowersby, Drew S.;Roberts, Rachel D.;Robson, Michael H.;Lewis, L. Kevin
蒙脱石等蒙脱石粘土与多种生物分子形成复合体，包括核酸DNA和RNA。以前对粘土吸附DNA的研究大多依赖于通过离心法将游离核酸从结合的络合物中分离出来后的分光光度分析。在目前的工作中，我们证明，由于(A)蒙脱石的不完全沉积和(B)剩余粘土在260 nm处的强吸收，这样的研究产生了一致的误差。粘土的沉降效率强烈依赖于阳离子浓度(Na+或Mg2+)和原始悬浮液的分散程度。建立了一种改进的粘土：DNA吸附实验，并利用该方法评价了金属反离子对单链DNA与蒙脱石结合的影响。X-射线衍射首次表明，形成的插层结构与平行于粘土表面的DNA链的取向一致。观察到的走廊间距被发现与利用原子建模技术计算的值非常接近。
Smectite clays such as montmorillonite form complexes with a variety of biomolecules, including the nucleic acids DNA and RNA. Most previous studies of DNA adsorption onto clay have relied upon spectrophotometric analysis after separation of free nucleic acids from bound complexes by centrifugation. In the current work we demonstrate that such studies produce a consistent error due to (a) incomplete sedimentation of montmorillonite and (b) strong absorbance of the remaining clay at 260 nm. Clay sedimentation efficiency was strongly dependent upon cation concentration (Na+ or Mg2+) and on the level of dispersion of the original suspension. An improved clay:DNA adsorption assay was developed and utilized to assess the impact of metal counterions on binding of single-stranded DNA to montmorillonite. X-ray diffraction demonstrated, for the first time, formation of intercalated structures consistent with orientation of the DNA strands parallel to the clay surface. Observed gallery spacings were found to closely match values calculated utilizing atomistic modeling techniques.
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