Sequence- and base-specific delivery of nitric oxide to cytidine and 5-methylcytidine leading to efficient deamination
Sequence- and base-specific delivery of nitric oxide to cytidine and 5-methylcytidine leading to efficient deamination
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DOI：
10.1021/ja0498888
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发表时间：
2004-07-28
影响因子：
15
通讯作者：
Sasaki, S
中科院分区：
文献类型：
--
作者：
Ali, M;Alam, R;Sasaki, S
一氧化氮（NO）是一种重要的内源性调节分子，亚硝基硫醇在NO的储存、运输和传递中起着重要作用。基于其在体内产生NO的能力，s -亚硝基硫醇可以用作治疗药物。在这项研究中，我们开发了一种创新的方法，用于序列和碱基特异性递送NO到DNA的特定位点，然后进行特异性脱氨。我们设计了一个从s -亚硝基硫鸟嘌呤到胞嘧啶的氨基互变异构体的NO转移反应。用s -亚硝基-n -乙酰青霉胺（SNAP）对含硫鸟苷的odn1进行亚硝化反应生成ODN2。利用odn2及其互补的odn3在靶位点具有dC或dmC，进行了链间NO转移反应，观察到快速的NO转移反应。相比之下，odn3在靶位点上含有dT、dA或dG， odn5 -7在非靶位点上含有dC，都没有观察到转移反应。在对no转移ODN4脱氨产物的分析中，发现dmC到dT的转化率高达42%，dmC重氮酸盐为13%。总之，我们已经证明了一种创新的方法，即序列和碱基特异性递送一氧化氮到胞苷和5-甲基胞苷。与传统方法相比，本研究显示的脱氨后NO转移的选择性和效率极高。
Nitric oxide (NO) is an important endogenous regulatory molecule, andS-nitrosothiols are believed to play a significant role in NO storage, transport, and delivery. On the basis of their ability to generate NO in vivo,S-nitrosothiols can be used as therapeutic drugs. In this study, we have developed an innovative method for sequence- and base-specific delivery of NO to a specific site of DNA followed by specific deamination. We designed a NO transfer reaction fromS-nitroso thioguanine to an imino tautomer of cytosine. Nitrosation of the thioguanosine-containing ODN1was carried out withS-nitroso-N-acetylpenicillamine (SNAP) to produce ODN2. An interstrand NO transfer reaction was performed using ODN2and its complementary ODN3having dC or dmC at the target site, and a rapid NO transfer reaction was observed. In contrast, a transfer reaction was not observed either with ODN3having dT, dA, or dG at the target site or with ODN 5-7 having dC at a nontarget site. In the analysis of deaminated products of the NO-transferred ODN4, it was found that the transformation ratio from dmC to dT was as high as 42% together with the dmC-diazoate (13%). In conclusion, we have demonstrated the innovative method of sequence- and base-specific delivery of nitric oxide to cytidine and 5-methylcytidine. The selectivity and efficiency of NO transfer followed by deamination exhibited in this study are extremely high compared to those of the conventional methods.