Medicinal Chemistry, Otto Loewi Research Center, Medical University of Graz, Graz, Austria.
BioTechMed-Graz, Graz, Austria.
Methods Mol Biol. 2025;2871:85-98. doi: 10.1007/978-1-0716-4217-7_8.
FOXOs are highly dynamic transcription factors consisting of one conserved DNA-binding domain (forkhead domain) as well as intrinsically disordered regions (IDRs) at the N- and C-termini. These IDRs are essential and regulate transcriptional activity of FOXOs by serving as interaction platform for cofactors. Furthermore, the IDRs are involved in intra- and intermolecular homeotypic and heterotypic interactions between FOXOs and in turn mediate FOXO auto-inhibition and condensate formation. Here, we describe generalizable methods to study the structure and dynamics of FOXO proteins in vitro using a combination of biophysical techniques, in particular nuclear magnetic resonance spectroscopy. These methods can serve as a blueprint for the investigation of other IDR-containing transcription factors.
FOXOs 是高度动态的转录因子，由一个保守的 DNA 结合结构域（叉头结构域）以及 N-和 C-末端的固有无序区域（IDR）组成。这些 IDR 是必不可少的，通过作为辅助因子的相互作用平台来调节 FOXO 的转录活性。此外，IDR 参与 FOXO 之间的分子内和分子间同源和异源相互作用，并反过来介导 FOXO 的自动抑制和凝聚体形成。在这里，我们描述了使用多种生物物理技术（特别是核磁共振波谱学）在体外研究 FOXO 蛋白结构和动力学的可推广方法。这些方法可以作为研究其他含有 IDR 的转录因子的蓝图。
