Biochemical and Biophysical Methods for Analysis of Poly(ADP-Ribose) Polymerase 1 and Its Interactions with Chromatin.
Biochemical and Biophysical Methods for Analysis of Poly(ADP-Ribose) Polymerase 1 and Its Interactions with Chromatin.
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DOI：
10.1007/978-1-4939-6993-7_16
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Luger K
中科院分区：
文献类型：
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作者：
Chassé MH;Muthurajan UM;Clark NJ;Kramer MA;Chakravarthy S;Irving T;Luger K
聚（ADP-核糖）聚合酶 I (PARP-1) 是 DNA 损伤修复的第一响应者，并参与基因表达的调节。 PARP-1 与染色质和 DNA 的相互作用很复杂，并且涉及至少两种不同的相互作用模式。在其酶促非活性状态下，PARP-1 结合天然染色质的亲和力与结合游离 DNA 末端的亲和力相似。 PARP-1 的自动修饰在不同程度上影响与染色质和 DNA 的相互作用。在这里，我们描述了一系列生物化学和生物物理技术来量化和剖析 PARP-1 与其各种底物的不同结合模式。此处列出的技术可对不同 PARP 构建体（非活性和自动修饰）与染色质和 DNA 损伤模型的相互作用进行高通量和定量测量。
Poly (ADP-Ribose) Polymerase I (PARP-1) is a first responder to DNA damage repair and participates in the regulation of gene expression. The interaction of PARP-1 with chromatin and DNA is complex and involves at least two different modes of interaction. In its enzymatically inactive state, PARP-1 binds native chromatin with similar affinity as it binds free DNA ends. Automodification of PARP-1 affects interaction with chromatin and DNA to different extents. Here we describe a series of biochemical and biophysical techniques to quantify and dissect the different binding modes of PARP-1 with its various substrates. The techniques listed here allow for high throughput and quantitative measurements of the interaction of different PARP constructs (inactive and automodified) with chromatin and DNA damage models.