Structures of oxaliplatin-oligonucleotide adducts from DNA.
Structures of oxaliplatin-oligonucleotide adducts from DNA.
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DOI：
10.1002/jms.3080
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发表时间：
2012-10
期刊：
影响因子：
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通讯作者：
S. Mowaka;Matthias Ziehe;D. Mohamed;U. Hochkirch;J. Thomale;M. Linscheid
中科院分区：
文献类型：
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作者：
S. Mowaka;Matthias Ziehe;D. Mohamed;U. Hochkirch;J. Thomale;M. Linscheid
奥沙利铂，[（1 R，2 R）-环己烷-1，2-二胺]（乙二酸-O，O '）铂（II）显示出对结肠直肠癌的极大有效性。尽管奥沙利铂的作用方式尚未明确，但普遍认为奥沙利铂与DNA的结合可阻止DNA合成并改变蛋白质与DNA的结合。为了阐明修饰的DNA-蛋白质相互作用，从而理解导致细胞误解DNA信息和细胞凋亡的机制，我们利用高效液相色谱/电喷雾串联质谱法，在寡聚体水平上确定了奥沙利铂-DNA链内和链间加合物的优先结合位点和动力学（HPLC/ESI-MS/MS）和HPLC/电感耦合等离子体-MS用于定量研究。我们使用benzonase、碱性磷酸酶和核酸酶S1的组合进行消化。这种消化过程允许铂化寡聚核苷酸和更复杂的链间加合物的研究。消化产物主要通过色谱法分离，并使用HPLC/ESI-离子阱MS/MS实验进行表征。我们可以证明鸟嘌呤和腺嘌呤的加合物是动态的，也就是说，它们的比例在几天内都在变化。此外，所得的加合物提供了证据的消化酶的作用，并表明，在寡聚体水平的加合物光谱是不同的，在通常研究的二核苷酸水平。
Oxaliplatin, [(1R,2R)-cyclohexane-1,2-diamine](ethanedioato-O,O')platinum(II) shows a great efficiency against colorectal cancer. Although the mode of action of oxaliplatin is not yet understood, it is commonly accepted that binding of oxaliplatin to DNA prevents DNA synthesis and alters protein to DNA binding. In order to elucidate the modified DNA-protein interaction and thus to understand the mechanisms leading to cellular misinterpretation of DNA information and apoptosis, we have identified the preferential binding sites and the dynamics of the oxaliplatin-DNA intrastrand and interstrand adducts at the oligomer level using high-performance liquid chromatography/electrospray ionization-tandem mass spectrometry (HPLC/ESI-MS/MS) and HPLC/inductively coupled plasma-MS for quantitative studies. We used a combination of benzonase, alkaline phosphatase and Nuclease S1 for digestion. This digestion procedure allows the study of platinated oligomeric nucleotides and more complex interstrand adducts. The digestion products were mostly chromatographically separated and characterized using HPLC/ESI-ion trap MS/MS experiments. We could show that the adducts to guanine and adenine are quite dynamic; that is, the ratios are changing for several days. In addition, the resulting adducts provide evidence for the action of the digesting enzymes and indicate that the adduct spectrum at the oligomeric level is different to that at the commonly studies dinucleotide level.