Beall Gary W, Sowersby Drew S, Roberts Rachel D, Robson Michael H, Lewis L Kevin
Department of Chemistry and Biochemistry, Texas State University, 601 University Drive, San Marcos, Texas 78666, USA.
Biomacromolecules. 2009 Jan 12;10(1):105-12. doi: 10.1021/bm800970v.
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 on spectrophotometric analysis after the separation of free nucleic acids from bound complexes by centrifugation. In the current work, we demonstrate that such studies produce a consistent error because of (a) incomplete sedimentation of montmorillonite and (b) strong absorbance of the remaining clay at 260 nm. Clay sedimentation efficiency was strongly dependent on 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 the binding of single-stranded DNA to montmorillonite. X-ray diffraction demonstrated, for the first time, the formation of intercalated structures consistent with orientation of the DNA strands parallel to the clay surface. Observed gallery spacings were found to match values calculated using atomistic modeling techniques closely.
蒙脱石等蒙脱石类粘土能与多种生物分子形成复合物,包括核酸DNA和RNA。此前大多数关于DNA吸附到粘土上的研究都依赖于通过离心将游离核酸与结合复合物分离后进行分光光度分析。在当前工作中,我们证明此类研究存在一致的误差,原因在于:(a)蒙脱石沉降不完全;(b)剩余粘土在260nm处有强吸收。粘土沉降效率强烈依赖于阳离子浓度(Na+或Mg2+)以及原始悬浮液的分散程度。我们开发并利用了一种改进的粘土-DNA吸附测定法来评估金属抗衡离子对单链DNA与蒙脱石结合的影响。X射线衍射首次证明形成了与DNA链平行于粘土表面排列一致的插层结构。观察到的层间距与使用原子模型技术计算的值紧密匹配。