Methidiumpropyl-EDTA.Fe(II) and DNase I footprinting report different small molecule binding site sizes on DNA.
Methidiumpropyl-EDTA.Fe(II) and DNase I footprinting report different small molecule binding site sizes on DNA.
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DOI：
10.1093/nar/11.16.5555
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发表时间：
1983-08
影响因子：
14.9
通讯作者：
M. V. Van Dyke;P. Dervan
中科院分区：
文献类型：
--
作者：
M. V. Van Dyke;P. Dervan
DNase I 和 MPE.Fe (II) 足迹分析均采用配体保护的 DNA 限制性片段的部分切割和 Maxam-Gilbert 测序凝胶分析方法。一种方法利用酶 DNase I 作为 DNA 切割剂，而另一种方法则采用合成分子甲锭-丙基-EDTA (MPE)。对于放线菌素 D、色霉素 A3 和偏端霉素 A，DNase I 足迹法报告的结合位点大小比 MPE.Fe (II) 更大。 DNase I 足迹似乎对弱结合位点更敏感。 MPE.Fe (II) 足迹法在确定 DNA 上小分子结合位点的实际大小和位置方面似乎更准确，特别是在 DNA 上多个小分子间隔很近的情况下。 MPE.Fe (II) 和 DNase I 报告操纵子 DNA 上 lac 阻遏蛋白的序列和结合位点大小相同。
DNase I and MPE.Fe (II) footprinting both employ partial cleavage of ligand-protected DNA restriction fragments and Maxam-Gilbert sequencing gel methods of analysis. One method utilizes the enzyme, DNase I, as the DNA cleaving agent while the other employs the synthetic molecule, methidium-propyl-EDTA (MPE). For actinomycin D, chromomycin A3 and distamycin A, DNase I footprinting reports larger binding site sizes than MPE.Fe (II). DNase I footprinting appears more sensitive for weakly bound sites. MPE.Fe (II) footprinting appears more accurate in determining the actual size and location of the binding sites for small molecules on DNA, especially in cases where several small molecules are closely spaced on the DNA. MPE.Fe (II) and DNase I report the same sequence and binding site size for lac repressor protein on operator DNA.