Ihara Toshihiro, Ishii Tatsuaki, Jyo Akinori
Department of Applied Chemistry and Biochemistry, Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan.
Nucleic Acids Symp Ser (Oxf). 2009(53):19-20. doi: 10.1093/nass/nrp010.
When designing ligands for specific sequences in DNA duplexes, triple helix formation is a useful recognition motif, because base triplet formation is based on the simple rule of complementary Hoogsteen hydrogen bonding, CG.C(+) and TA.T. However the triplexes containing CG.C(+) triplets form only in a weak acidic solution, because cytosines in third strand need to be protonated to satisfy its complementarity to CG base-pairs. A simple and easy method to stabilize the DNA triplex using Ag(+) was reported. A silver ion displaces the N3 proton of cytosine in Hoogsteen base-pairing to form a base triplet, CG.CAg(+). By the addition of an equimolar amount of Ag(+), the third strand 15 mer sequence containing five cytosines was stabilized by ca. 30 degrees C in melting temperature at pH 7. The triplex structure was stable even under weak basic conditions.
在设计用于DNA双链体特定序列的配体时,三链螺旋形成是一种有用的识别基序,因为碱基三联体的形成基于互补的Hoogsteen氢键的简单规则,即CG.C(+)和TA.T。然而,含有CG.C(+)三联体的三链体仅在弱酸性溶液中形成,因为第三链中的胞嘧啶需要质子化才能满足其与CG碱基对的互补性。据报道,有一种使用Ag(+)稳定DNA三链体的简单易行的方法。银离子取代Hoogsteen碱基配对中胞嘧啶的N3质子以形成碱基三联体CG.CAg(+)。通过加入等摩尔量的Ag(+),含有五个胞嘧啶的15聚体第三链序列在pH 7时的解链温度下稳定了约30℃。即使在弱碱性条件下,三链体结构也是稳定的。