Mapping weak and transient UBL:receptor interactions using tailored photo-crosslinking chemistry

蛋白质的翻译后修饰(PTM)是几乎所有细胞活动的关键。这包括DNA损伤反应(DDR)，这是一个信号网络，通过预防癌症和神经退行性病变等与年龄相关的疾病，对促进健康老龄化至关重要。特别是，泛素样蛋白(UBL)等PTM最近已成为各种DNA修复途径的关键调节因子1。因此，阐明与UBL结合的生化后果对于理解这些PTM标记的功能相关性至关重要。因此，已经出现了大量的努力来识别和表征下游的UBL结合蛋白--也被称为受体--它们解释UBL标记并将其翻译成精确的细胞活动。然而，Ubl：受体相互作用的系统鉴定是具有挑战性的，因为它们大多是弱的和瞬时的。为了应对这一挑战，我们的目标是将一系列化学生物学工具与遗传密码扩展相结合，以系统地绘制管理UBL网络的微弱和瞬时相互作用，这些网络最近已成为DDR的重要翻译后修饰因子。

Post-translational modification (PTM) of proteins is key to virtually all cellular activities. This includes the DNA damage response (DDR), a signalling network critical for facilitating healthy ageing by preventing age-related diseases such as cancer and neurodegenerative pathologies. In particular, PTMs such as ubiquitin-like proteins (UBLs) have recently emerged as critical regulators of a variety of DNA repair pathways1. Elucidating the biochemical consequences of conjugation with UBLs is therefore critical for understanding the functional relevance of these PTM marks. Intense efforts have therefore emerged to identify and characterise the downstream UBL-binding proteins - also called receptors - that interpret and translate UBL marks into precise cellular activities. However, systematic identification of UBL:receptor interactions is challenging due to their mostly weak and transient nature. To address this challenge we aim to integrate a range of chemical biology tools with genetic code expansion to systematically map weak and transient interactions governing UBL networks that have recently emerged as important posttranslational modifiers of the DDR.