PDF(Pubmed)
Abstract：
Over the past decade, the connection between APOBEC3 cytosine deaminases and cancer mutagenesis has become increasingly apparent. This growing awareness has created a need for biochemical tools that can be used to identify and characterize potential inhibitors of this enzyme family. In response to this challenge, we have developed a Real-time APOBEC3-mediated DNA Deamination assay. This assay offers a single-step set-up and real-time fluorescent read-out, and it is capable of providing insights into enzyme kinetics. The assay also offers a high-sensitivity and easily scalable method for identifying APOBEC3 inhibitors. This assay serves as a crucial addition to the existing APOBEC3 biochemical and cellular toolkit and possesses the versatility to be readily adapted into a high-throughput format for inhibitor discovery.
摘要：
在过去的十年里，APOBEC3胞嘧啶脱氨酶与癌症诱变之间的联系越来越明显。这种日益增长的意识产生了对可用于鉴定和表征该酶家族的潜在抑制剂的生化工具的需求。为了应对这一挑战，我们开发了一种实时APOBEC3介导的DNA脱氨(RADD)检测方法。该测定提供了单步设置和实时荧光读出，它能够提供对酶动力学的见解，并提供高灵敏度和易于扩展的方法来鉴定APOBEC3抑制剂。该测定作为对现有APOBEC3生物化学和细胞工具包的重要补充,并且具有多功能性,易于适应于发现抑制剂的高通量形式。
