基于分子信标对hOGG1活性的定量分析

Quantitative Analysis of hOGG1 Activity Based on Molecular Beacon

设计了一种含有8-oxoG碱基的新型分子信标,结合酶促反应发展了一种非同位素标记的人8-oxoG-鸟嘌呤糖苷酶1(hOGG1)的活性分析新方法,检出限可达0.0125 U/mL.此外,该方法还可用于快速考察金属离子对酶促反应的影响和肿瘤细胞中hOGG1活性水平的定量检测.实验结果表明,该方法简单、灵敏,有望用于肿瘤样品中hOGG1活性的高通量分析和hOGG1抑制剂的筛选.

Human 8-oxo-guanine DNA glycosylase 1（hOGG1）,which mainly catalyzes the excision of 8-oxoG and cleaves the AP site from DNA to sustain the genome integrity,plays an important role in many biological events including tumor development.However,available methods of hOGG1 assay based on gel electrophoresis are complicate,laborious and discontinuous.Here,we report a new method to analyze hOGG1 activity using molecular beacon containing 8-oxoG coupled with enzyme-linkage reactions,the detection limitation of this method is 0.0125 U/mL.Furthermore,the method was applied for quantification of hOGG1 activity in tumor cells and investigating the effect of metal ions on the reaction.The results demonstrate that the easy,simple and non-radioactive method has the potential to screen suitable inhibitors for hOGG1 and can be applied for high-throughput analysis of hOGG1 activity in tumor samples.