FLUORESCENCE ANISOTROPY ASSAYS IMPLICATE PROTEIN-PROTEIN INTERACTIONS IN REGULATING TRP REPRESSOR DNA-BINDING
FLUORESCENCE ANISOTROPY ASSAYS IMPLICATE PROTEIN-PROTEIN INTERACTIONS IN REGULATING TRP REPRESSOR DNA-BINDING
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DOI：
10.1021/bi00081a021
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发表时间：
1993-08-03
期刊：
影响因子：
2.9
通讯作者：
ROYER, CA
中科院分区：
文献类型：
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作者：
LETILLY, V;ROYER, CA
蛋白质和核酸之间相互作用的研究是理解基因表达控制的核心。荧光各向异性已被用来测量，在溶液中，细菌阻遏蛋白，色氨酸阻遏蛋白（TR），含有其目标操纵基因序列之一的荧光标记的寡核苷酸的平衡结合概况。改变辅阻遏物、操纵基因DNA和蛋白质浓度的影响的研究表明TR寡聚体参与DNA结合的调节。这些研究还表明，相对简单的荧光各向异性技术可以应用于蛋白质和核酸之间的相互作用的研究。荧光技术具有足够的灵敏度，在大多数情况下可以取代放射性检测方法。此外，由于它是一种基于溶液的方法，它提供了蛋白质-核酸平衡的真实平衡测量，并且可以容易地评估溶液条件（例如作为盐和配体浓度、pH和温度）变化的影响。数据采集相对简单和快速，数据质量足以进行复杂系统的详细热力学分析。鉴于这些属性，荧光各向异性将在遗传调控领域中找到多种应用。
The study of interactions between proteins and nucleic acids is central to the understanding of the control of genetic expression. Fluorescence anisotropy has been used to measure, in solution, the equilibrium binding profiles of a bacterial repressor protein, the tryptophan repressor (TR), to a fluorescently labeled oligonucleotide containing one of its target operator sequences. Investigation of the effects of changing concentrations of corepressor, operator DNA, and protein implicate TR oligomers in the regulation of DNA binding. These studies also demonstrate that the relatively straightforward technique of fluorescence anisotropy can be applied to the study of the interactions between proteins and nucleic acids. The fluorescence technique exhibits sufficient sensitivity to replace radioactive methods of detection in most cases. In addition, since it is a solution-based methodology, it offers a true equilibrium measure of the protein-nucleic acid equilibria, and the effects of changes in solution conditions such as salt and ligand concentration, pH, and temperature can be readily evaluated. Data acquisition is relatively simple and rapid, and the data are of sufficient quality for detailed thermodynamic analyses of complex systems. Given these attributes, fluorescence anisotropy will find multiple applications in the area of genetic regulation.