Kageyama Tomoe, Sato Yusuke, Nishizawa Seiichi, Teramae Norio
Department of Chemistry, Graduate School of Science, Tohoku university, and CREST, Japan Science and Technology Agency (JST), Aoba-ku, Sendai 980-8578, Japan.
Nucleic Acids Symp Ser (Oxf). 2008(52):119-20. doi: 10.1093/nass/nrn061.
By using lumichrome (Lch) as a masking ligand, we successfully control the binding selectivity of 2-amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND) when binding to nucleobases in AP site-containing DNA duplexes (5'-TCT GCG TCC AGX GCA ACG CAC AC-3'/3'-AGA CGC AGG TCN CGT TGC GTG TG-5', X = AP site; Spacer C3, N = C or T). In solutions buffered to pH 7.0 (I = 0.11 M, at 5 degrees C), ATMND binds to cytosine and thymine with a comparable binding affinity (K(d) / nM: C: 7.7, T: 15). By contrast, in the presence of Lch, ATMND shows a clear binding selectivity for cytosine over thymine (K(d)/nM: C: 17, T: 204). Such competitive binding events are discussed with a view towards development of ligand-based fluorescence assay for single-nucleotide polymorphisms (SNPs) typing.
通过使用核黄素（Lch）作为掩蔽配体，我们成功地控制了2-氨基-5,6,7-三甲基-1,8-萘啶（ATMND）与含脱嘌呤嘧啶（AP）位点的DNA双链体（5'-TCT GCG TCC AGX GCA ACG CAC AC-3'/3'-AGA CGC AGG TCN CGT TGC GTG TG-5'，X = AP位点；间隔物C3，N = C或T）中的核碱基结合时的结合选择性。在缓冲至pH 7.0（I = 0.11 M，5℃）的溶液中，ATMND以相当的结合亲和力与胞嘧啶和胸腺嘧啶结合（解离常数K(d)/nM：C：7.7，T：15）。相比之下，在Lch存在下，ATMND对胞嘧啶显示出比对胸腺嘧啶更明显的结合选择性（K(d)/nM：C：17，T：204）。针对基于配体的单核苷酸多态性（SNP）分型荧光测定法的开发，对这种竞争性结合事件进行了讨论。
