HYDROXYL RADICAL FOOTPRINTING - HIGH-RESOLUTION INFORMATION ABOUT DNA PROTEIN CONTACTS AND APPLICATION TO LAMBDA-REPRESSOR AND CRO PROTEIN
HYDROXYL RADICAL FOOTPRINTING - HIGH-RESOLUTION INFORMATION ABOUT DNA PROTEIN CONTACTS AND APPLICATION TO LAMBDA-REPRESSOR AND CRO PROTEIN
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DOI：
10.1073/pnas.83.15.5469
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发表时间：
1986-08-01
影响因子：
11.1
通讯作者：
DOMBROSKI, BA
中科院分区：
文献类型：
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作者：
TULLIUS, TD;DOMBROSKI, BA
已经开发出一种方法来制作与DNA结合的蛋白质的“足迹”。羟基自由基，由铁（II）还原过氧化氢产生，是用于切割DNA的试剂。羟基自由基破坏DNA的主链，几乎没有序列依赖性，因此可以监测所有主链位置与蛋白质的接触。除了定义与蛋白质接触的DNA序列外，羟基自由基足迹还包含有关DNA-蛋白质复合物的结构信息。例如，噬菌体的羟基自由基足迹。阻遏物和Cro蛋白直接表明这些蛋白质仅结合于DNA螺旋的一侧。其他联系人. darda.阻遏物和Cro蛋白与DNA，没有观察到的其他化学足迹法的羟基自由基足迹揭示。
A method has been developed for making "footprints" of proteins bound to DNA. The hydroxyl radical, generated by reduction of hydrogen peroxide by iron(II), is the reagent used to cut the DNA. Hydroxyl radical breaks the backbone of DNA with almost no sequence dependence, so all backbone positions may be monitored for contact with protein. In addition to defining the DNA sequence in contact with the protein, hydroxyl radical footprints embody structural information about the DNA-protein complex. For example, hydroxyl radical footprints of the bacteriophage .lambda. repressor and Cro protein show directly that these proteins are bound to only one side of the DNA helix. Additional contacts of .lambda. repressor and Cro protein with DNA, not observed by other chemical footprinting methods, are revealed by hydroxyl radical footprinting.