Drug design for transcription factors

我们开发了新的技术来分析DNA结合蛋白的功能以及蛋白质与其他小分子之间的相互作用。正在开发的新方法包括检测蛋白质DNA结合特异性的双链DNA芯片和蛋白质循环流动核磁共振系统，该系统将允许更快地筛选与蛋白质相互作用的化合物，同时使用更少的被筛选蛋白质。通过使用新开发的双链DNA芯片，我们已经快速检测了结合双链端粒DNA的人TRF1的序列特异性。在玻片上切下多条不同序列的双链DNA，然后在玻片上检测到hTRF1。荧光标记DNA的强度反映了hTRF1的序列特异性。结合hTRF1的特定DNA序列的每个强度与用Biacore检测的每个DNA序列的hTRF1的结合解离常数有很好的相关性。使用Biacore分析hTRF1对7种不同序列的结合常数需要7天，而使用我们新的双链DNA芯片需要半个小时。

We have developed new technologies to analyze DNA-binding protein function and the interaction between proteins and other small molecules. New approach under development include a double-stranded DNA chip to detect DNA-binding specificities of proteins and a Protein Recycling Flow NMR system that will allow for faster screening of compounds that interact with proteins, while using much smaller amounts of the protein being screened.By using the newly developed double-stranded DNA chip we have detected rapidly the sequence specificity of human TRF1 that binds double stranded telomeric DNA. Many double-stranded DNA with different sequences were chipped on a glass slide and then hTRF1 was spotted on the glass slide. The intensities of the fluorescence labeled DNA indicate the sequence specificities of hTRF1. The each intensity of a specific DNA sequence bound hTRF1 is well correlated to the binding dissociation constant of hTRF1 for each DNA sequence examined by using BIACORE. By using BIACORE it takes seven days to analyze the binding constants of hTRF1 for seven different sequences, while by using our new double-stranded DNA chip it takes half an hour.