Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, California 90089, United States.
Department of Molecular Medicine, The Scripps Research Institute, Jupiter, Florida 33458, United States.
ACS Chem Biol. 2021 Feb 19;16(2):389-396. doi: 10.1021/acschembio.0c00937. Epub 2021 Feb 1.
Protein poly-ADP-ribosylation (PARylation) is a heterogeneous and dynamic post-translational modification regulated by various writers, readers, and erasers. It participates in a variety of biological events and is involved in many human diseases. Currently, tools and technologies have yet to be developed for unambiguously defining readers and erasers of individual PARylated proteins or cognate PARylated proteins for known readers and erasers. Here, we report the generation of a bifunctional nicotinamide adenine dinucleotide (NAD) characterized by diazirine-modified adenine and clickable ribose. By serving as an excellent substrate for poly-ADP-ribose polymerase 1 (PARP1)-catalyzed PARylation, the generated bifunctional NAD enables photo-cross-linking and enrichment of PARylation-dependent interacting proteins for proteomic identification. This bifunctional NAD provides an important tool for mapping cellular interaction networks centered on protein PARylation, which are essential for elucidating the roles of PARylation-based signals or activities in physiological and pathophysiological processes.
蛋白质聚 ADP-核糖基化（PARylation）是一种由各种writers、readers 和 erasers 调控的异质和动态的翻译后修饰。它参与了多种生物事件，并与许多人类疾病有关。目前，还没有开发出用于明确定义已知 readers 和 erasers 的单个 PARylated 蛋白或同源 PARylated 蛋白的 readers 和 erasers 的工具和技术。在这里，我们报告了一种双功能烟酰胺腺嘌呤二核苷酸（NAD）的生成，其特征是腺嘌呤经过重氮修饰和可点击的核糖。作为聚 ADP-核糖聚合酶 1（PARP1）催化的 PARylation 的极好底物，生成的双功能 NAD 能够进行光交联和 PARylation 依赖性相互作用蛋白的富集，用于蛋白质组学鉴定。这种双功能 NAD 为绘制以蛋白质 PARylation 为中心的细胞相互作用网络提供了重要工具，这对于阐明 PARylation 基信号或活性在生理和病理生理过程中的作用至关重要。
