Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
Bioconjug Chem. 2009 Aug 19;20(8):1643-9. doi: 10.1021/bc900200c. Epub 2009 Jul 8.
Two types of 5'-β-CyD-DNA conjugates were synthesized using two different strategies and were hybridized with cDNA conjugates bearing various possible guest compounds at the 3' ends. One of the β-CyD conjugates was synthesized on the basis of solid-phase postmodification of DNA with a monoamino-β-CyD derivative on a synthesis column for automated DNA synthesis. The other β-CyD conjugate was synthesized by the solution-phase coupling of DNA with a monomercapto-β-CyD derivative using a heterobifunctional cross-coupling reagent. When these 5'-β-CyD-DNA conjugates were hybridized with cDNA conjugates bearing 1-adamantaneacetic acid at the 3' ends, significant stabilization of the duplexes was observed as compared with the control duplex without β-CyD. Duplexes of 5'-β-CyD-DNA conjugates and complementary 3'-dansyl-glycine-DNA conjugates were also moderately stabilized. Thermodynamic measurements revealed that the host-guest inclusion interactions between β-CyD and 1-adamantaneacetic acid or dansyl-glycine are roughly as strong as those found in bulk solution even if they are tethered to the ends of the DNA.
两种 5'-β-CyD-DNA 缀合物是使用两种不同的策略合成的，并与在 3' 末端带有各种可能的客体化合物的 cDNA 缀合物杂交。其中一种 β-CyD 缀合物是基于在合成柱上用单氨基-β-CyD 衍生物对 DNA 进行固相后修饰合成的，用于自动化 DNA 合成。另一种 β-CyD 缀合物是通过使用杂双功能偶联试剂将 DNA 与单巯基-β-CyD 衍生物在溶液相中偶联合成的。当这些 5'-β-CyD-DNA 缀合物与在 3' 末端带有 1-金刚烷乙酸的 cDNA 缀合物杂交时，与没有 β-CyD 的对照双链体相比，观察到双链体有显著的稳定化。5'-β-CyD-DNA 缀合物和互补的 3'-丹磺酰基甘氨酸-DNA 缀合物的双链体也得到适度稳定。热力学测量表明，即使 β-CyD 与 1-金刚烷乙酸或丹磺酰基甘氨酸的主体 - 客体包合相互作用被束缚在 DNA 的末端，它们与在体相溶液中发现的相互作用大致相同。
