College of Life Science, Guizhou University, Guiyang 550025, China.
Bioorg Med Chem Lett. 2012 Jul 1;22(13):4238-41. doi: 10.1016/j.bmcl.2012.05.044. Epub 2012 May 17.
8-17 DNAzyme is characterized by its recurrence in different in vitro selections and versatile cleavage sites, leading to extensive studies on its structural properties and applications. We evaluated the purine residues (A6, G7, G11, A12, G14, and A15) in the catalytic core of 8-17 DNAzyme of their five-membered ring moiety with purine analogs 1-5 to have an insight into the conservation of the residues at the level of functional groups. The 7-nitrogen atom in the AGC loop was demonstrated to be strictly conserved for the cleavage reaction. But such modifications exerted favorable effect at G11 of the base-pair stem and A12 in the single-strand loop, directing toward more efficient DNAzymes. Even the most conserved G14 could tolerate such modifications. These results demonstrated that chemical modification on the functional groups is a feasible approach to gain an insight into the structural requirement in the catalytic reaction of DNAzymes. It also provided modification sites for introduction of signaling molecules used for mechanistic and folding studies of 8-17 DNAzyme.
8-17 DNA 酶的特点是在不同的体外选择和多种切割位点中重复出现,这导致了对其结构特性和应用的广泛研究。我们评估了催化核心中的嘌呤残基(A6、G7、G11、A12、G14 和 A15)在 8-17 DNA 酶的五元环部分的嘌呤类似物 1-5,以深入了解残基在功能基团水平上的保守性。AGC 环中的 7-氮原子被证明对切割反应是严格保守的。但是,这种修饰对碱基对茎中的 G11 和单链环中的 A12 产生了有利的影响,导致更有效的 DNA 酶。即使是最保守的 G14 也能耐受这种修饰。这些结果表明,对功能基团进行化学修饰是深入了解 DNA 酶催化反应结构要求的一种可行方法。它还为引入用于 8-17 DNA 酶的机制和折叠研究的信号分子提供了修饰位点。