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），这是一种通过预防癌症和神经退行性病变等与年龄相关的疾病来促进健康老龄化的关键信号网络。特别是，PTM如泛素样蛋白（UBLs）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缀合的生物化学后果对于理解这些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鉴定和鉴定下游UBL结合蛋白-也称为受体-将UBL标记解释和翻译成精确的细胞活性。然而，系统鉴定UBL：受体相互作用是具有挑战性的，因为它们大多是弱的和短暂的性质。为了应对这一挑战，我们的目标是整合一系列的化学生物学工具与遗传密码扩展，系统地映射弱和短暂的相互作用，最近出现的UBL网络，作为重要的翻译后修饰的DDR。

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.