Lv Wei, Li Na, Li Yuliang, Li Ye, Xia Andong
State Key Laboratory of Molecular Reaction Dynamics, Institute of Chemistry, Chinese Academy of Sciences, Beijing-100080, P. R. China.
J Am Chem Soc. 2006 Aug 9;128(31):10281-7. doi: 10.1021/ja063004y.
We present the detection of the shape-specific conformation of DNA based on the fluorescence resonance energy transfer (FRET) by using a novel flexible water-soluble cationic conjugated polymer (CCP). The flexible backbone of CCP has more conformational freedom with the potential to be responsive to analyte shape by electrostatic interaction between flexible CCP and negatively charged DNA. The analyte shape dependent recognition is accomplished by structural changes that compressed or extended the flexible CCP. The morphology-dependent spectral properties of the novel flexible polymer related to the analyte shapes are investigated in detail, where two types of chromophores, referred to as "isolated" segment and "packed" segment aggregates, within the flexible polymer are identified by means of ensemble and single molecule measurements upon binding with different geometric DNA. The change in fluorescence intensity upon binding with shape-specific DNA without obvious color shifts makes this novel flexible polymer a suitable CCP donor for FRET measurements. The results provide insights for understanding the spectral properties of flexible water-soluble CCP and CCP/DNA interaction related to the geometry of target analyte.
我们展示了通过使用一种新型柔性水溶性阳离子共轭聚合物(CCP),基于荧光共振能量转移(FRET)对DNA的形状特异性构象进行检测。CCP的柔性主链具有更大的构象自由度,有可能通过柔性CCP与带负电荷的DNA之间的静电相互作用对分析物形状做出响应。分析物形状依赖性识别是通过压缩或伸展柔性CCP的结构变化来实现的。详细研究了与分析物形状相关的新型柔性聚合物的形态依赖性光谱特性,其中通过与不同几何形状的DNA结合后的系综和单分子测量,识别出柔性聚合物内的两种发色团,即“孤立”段和“堆积”段聚集体。与形状特异性DNA结合时荧光强度的变化而无明显颜色变化,使得这种新型柔性聚合物成为用于FRET测量的合适CCP供体。这些结果为理解柔性水溶性CCP的光谱特性以及与目标分析物几何形状相关的CCP/DNA相互作用提供了见解。