Physik Department & Munich School of Bioengineering, Technische Universität München, Am Coulombwall 4a, 85784, Garching, Germany.
Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford, OX1 3QZ, UK.
Angew Chem Int Ed Engl. 2019 Jun 3;58(23):7662-7667. doi: 10.1002/anie.201901734. Epub 2019 Apr 29.
Protein-protein and protein-substrate interactions are critical to function and often depend on factors that are difficult to disentangle. Herein, a combined biochemical and biophysical approach, based on electrically switchable DNA biochips and single-molecule mass analysis, was used to characterize the DNA binding and protein oligomerization of the transcription factor, forkhead box protein P2 (FOXP2). FOXP2 contains domains commonly involved in nucleic-acid binding and protein oligomerization, such as a C H -zinc finger (ZF), and a leucine zipper (LZ), whose roles in FOXP2 remain largely unknown. We found that the LZ mediates FOXP2 dimerization via coiled-coil formation but also contributes to DNA binding. The ZF contributes to protein dimerization when the LZ coiled-coil is intact, but it is not involved in DNA binding. The forkhead domain (FHD) is the key driver of DNA binding. Our data contributes to understanding the mechanisms behind the transcriptional activity of FOXP2.
蛋白质-蛋白质和蛋白质-底物相互作用对功能至关重要，而这些相互作用通常依赖于难以区分的因素。在此，我们采用了一种组合的生化和生物物理方法，基于电可切换 DNA 生物芯片和单分子质量分析，来表征转录因子叉头框蛋白 P2（FOXP2）的 DNA 结合和蛋白质寡聚化。FOXP2 包含常见的参与核酸结合和蛋白质寡聚化的结构域，如 C H -锌指（ZF）和亮氨酸拉链（LZ），但其在 FOXP2 中的作用在很大程度上仍不清楚。我们发现，LZ 通过形成卷曲螺旋介导 FOXP2 二聚体，但也有助于 DNA 结合。当 LZ 卷曲螺旋完整时，ZF 有助于蛋白质二聚体的形成，但它不参与 DNA 结合。叉头结构域（FHD）是 DNA 结合的关键驱动因素。我们的数据有助于理解 FOXP2 转录活性背后的机制。
