Interaction of DNA binding domain of HNF‐3α with its transferrin enhancer DNA specific target site
Interaction of DNA binding domain of HNF‐3α with its transferrin enhancer DNA specific target site
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HNF-3α DNA 结合域与其转铁蛋白增强子 DNA 特异性靶位点的相互作用
DOI：
10.1016/0014-5793(95)00767-4
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发表时间：
1995
期刊：
影响因子：
3.5
通讯作者：
M. Zakin
中科院分区：
文献类型：
--
作者：
H. Terenzi;I. Petropoulos;C. Ellouze;Masayuki Takahashi;M. Zakin
转铁蛋白肝特异性基因增强子（transferrin hepatic gene enhancer，TFR）是一种复杂的分子构筑物，与肝脏富集的HNF-3 α转录因子和普遍存在的蛋白质相关，通过DNA-蛋白质和蛋白质-蛋白质相互作用维持。作为理解其组织和活性机制的第一步，我们通过不同的生物物理技术分析了HNF-3 α（HDBD）的DNA结合结构域与转铁蛋白增强子中存在的特定DNA片段的相互作用。使用表面等离子体共振测量这种相互作用的动力学常数。用圆二色谱和荧光光谱对HDBD与DNA的相互作用进行了表征。HDBD以高亲和力（Kd = 10 − 8M）与其特异性DNA位点结合。在靶DNA的序列修饰后，亲和力降低。尺寸排阻色谱法和结合化学计量法测定的荧光测量表明，蛋白质是以单体形式存在之前和之后与DNA的相互作用。蛋白质的二级结构在与特异性DNA结合后没有显著改变。相反，通过与HDBD的相互作用，DNA的结构变化似乎发生。
Transferrin hepato-specific gene enhancer, associated with the liver-enriched HNF-3α transcriptional factor and ubiquitous proteins, is a complex molecular edifice maintained through DNA-protein and protein-protein interactions. As a first step to understand the mechanisms responsible for its organization and activity, we have analyzed the interaction of the DNA binding domain of HNF-3α (HDBD) with a specific DNA segment present in the transferrin enhancer by different biophysical techniques. The kinetic constants of this interaction were measured using surface plasmon resonance. The HDBD-DNA interaction was also characterized by circular dichroism and fluorescence spectroscopy. HDBD binds to its specific DNA site with high affinity (Kd≊10−8M ). The affinity is reduced after sequence modification of the target DNA. Size exclusion chromatography and binding stoichiometry determined by fluorescence measurements indicate that the protein is present in a monomeric form before and after interaction with the DNA. The secondary structure of the protein was not significantly altered upon binding to specific DNA. By contrast, a structural change of DNA by interaction with HDBD seems to occur.
影响因子：
2.9
作者：
JOHNSON, BB;DAHL, KS;ZHURKIN, VB
通讯作者：
ZHURKIN, VB
影响因子：
10.5
作者：
LAI, E;PREZIOSO, VR;DARNELL, JE
通讯作者：
DARNELL, JE
影响因子：
10.5
作者：
LAI, E;PREZIOSO, VR;DARNELL, JE
通讯作者：
DARNELL, JE