Structure elucidation of DNA interstrand cross-link by a combination of nuclease P1 digestion with mass spectrometry
Structure elucidation of DNA interstrand cross-link by a combination of nuclease P1 digestion with mass spectrometry
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DOI：
10.1021/ac034683n
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发表时间：
2003-11-15
影响因子：
7.4
通讯作者：
Wang, YS
中科院分区：
文献类型：
--
作者：
Wang, YS;Wang, YS
DNA链间交联试剂是治疗癌症最有效的试剂之一。在这里，我们报告了一种结合核酸酶P1酶切/质谱法的结构解析双寡核苷酸（ODNs）含有链间交联。我们的研究结果表明，核酸酶P1酶切含有链间交联（ICL）的4,5'，8-三甲基补骨脂素或丝裂霉素C的双链ODN得到一个带有交联核碱基片段的四核苷酸。四核苷酸去质子化离子的产物离子谱提供了交联结构的信息。此外，含有丝裂霉素C链间交联的两种取向异构体的四核苷酸的产离子谱也不同。我们相信本文所描述的方法可以普遍适用于研究其他DNA ICLs的结构。
DNA interstrand cross-link reagents are among the most powerful agents for cancer treatment. Here we report a combined nuclease P1 digestion/mass spectrometry method for the structure elucidation of duplex oligode-oxynucleotides (ODNs) containing an interstrand cross-link. Our results demonstrate that nuclease P1 digestion of a double-stranded ODN containing an interstrand cross-link (ICL) of 4,5',8-trimethylpsoralen or mitomycin C gives a tetranucleotide bearing the cross-linked nucleobase moiety. Product ion spectra of the deprotonated ions of the tetranucleotides provide information about the structure of the cross-link. Furthermore, product-ion spectra of tetranucleotides containing two orientation isomers of mitomycin C interstrand cross-link are distinctive. We believe that the method described in this paper can be generally applicable for investigating the structures of other DNA ICLs.