Winnacker Malte, Breeger Sascha, Strasser Ralf, Carell Thomas
Ludwig-Maximilians University Munich, Center for Integrated Protein Science (CiPSM), Department for Chemistry and Biochemistry, Butenandtstrasse 5-13, Munich, Germany.
Chembiochem. 2009 Jan 5;10(1):109-18. doi: 10.1002/cbic.200800397.
An investigation of the precise interactions between damaged DNA and DNA repair enzymes is required in order to understand the lesion recognition step, which is one of the most fundamental processes in DNA repair. Most recently, photoaffinity labeling approaches have enabled the analysis of even transient protein-DNA interactions. Here we report the synthesis and evaluation of oligonucleotides that contain two photoaffinity "catcher moieties" next to incorporated DNA lesions. With these DNA constructs it is possible to analyze the interactions between DNA lesions and the appropriate repair enzymes. The probes labeled the repair protein efficiently enough to enable subsequent protein analysis by mass spectrometry.
为了理解损伤识别步骤（这是DNA修复中最基本的过程之一），需要对受损DNA与DNA修复酶之间的精确相互作用进行研究。最近，光亲和标记方法甚至能够分析瞬时的蛋白质-DNA相互作用。在此，我们报告了在掺入的DNA损伤旁边含有两个光亲和“捕获基团”的寡核苷酸的合成与评估。利用这些DNA构建体，可以分析DNA损伤与适当修复酶之间的相互作用。这些探针能够有效地标记修复蛋白，以便随后通过质谱法进行蛋白质分析。
