Regulation of transcriptional activation by protein disorder

标题：用于测量蛋白质-蛋白质和蛋白质-蛋白质的荧光板读数器的获得 DNA结合亲和力 总结 了解蛋白质-蛋白质和蛋白质-DNA结合是深入了解蛋白质的必要条件。 蛋白质功能及其与疾病的关系荧光偏振（FP）是一种出色的工具， 用于测量蛋白质-蛋白质和蛋白质DNA结合亲和力， 等温滴定量热法（ITC）。现代FP仪器可以测量结合亲和力 是否产生热量，并使用酶标仪进行高通量和快速检测。 我们申请资金用于Agilent BioTek Synergy H1酶标仪，以提高我们的能力， 在盐浓度范围内测量p53突变体和DNA之间的结合亲和力，其中 无法检测到来自ITC的热信号。这些实验的结果将提高我们的 了解p53 DNA结合特异性如何通过分子内相互作用调节 之间的反式激活和DNA结合域，这是一个主要的目标， 母公司奖励（R 01 GM 115556）。

Title: Acquisition of a fluorescent plate reader for measuring protein-protein and protein- DNA binding affinities Summary Understanding protein-protein and protein-DNA binding is required for a deep understanding of protein function and its relation to disease. Fluorescence polarization (FP) is an outstanding tool for measuring protein-protein and protein DNA binding affinities, and it is more flexible than isothermal titration calorimetry (ITC). Modern FP instruments can measure binding affinities whether heat is produced and use a microplate reader for high throughput and rapid detection. We request funds for the Agilent BioTek Synergy H1 microplate reader to improve our ability to measure binding affinity between p53 mutants and DNA over a range of salt concentrations where a heat signal from ITC cannot be detected. Results from these experiments will improve our understanding of how p53 DNA binding specificity is regulated by an intramolecular interaction between the transactivation and DNA binding domains, which is one of the main goals of the parent award (R01GM115556).