Abstract：
Covalent sensors to detect and capture aggregated proteome in stressed cells are rare. Herein, we construct a series of covalent fluorogenic sensors for aggregated proteins by structurally modulating GFP chromophore and arming it with an epoxide warhead. Among them, P2 probe selectively modifies aggregated proteins over folded ones and turns on fluorescence as evidenced by biochemical and mass spectrometry results. The coverage of this epoxide-based covalent chemistry is demonstrated using different types of aggregated proteins. Finally, the covalent fluorescent sensor P2 allows for direct visualization and capture of aggregated proteome in stressed cardiomyocytes and cardiac tissue samples from a cardio-oncology mouse model. The epoxide-based covalent sensor developed herein may become useful for future chemical proteomics analysis of aggregated proteins to dissect the mechanism underlying cardio-oncology.
摘要：
在应激细胞中检测和捕获聚集的蛋白质组的共价传感器是罕见的。在这里，我们通过在结构上调节GFP发色团并用环氧化物弹头装备它，为聚集的蛋白质构建了一系列共价荧光传感器。其中,P2探针在折叠的蛋白质上选择性地修饰聚集的蛋白质，并打开荧光，如生物化学和质谱结果所证明的。使用不同类型的聚集蛋白质证明了这种基于环氧化物的共价化学的覆盖范围。最后,共价荧光传感器P2允许直接可视化和捕获来自心脏肿瘤小鼠模型的应激心肌细胞和心脏组织样品中的聚集蛋白质组。本文开发的基于环氧化物的共价传感器可用于聚集蛋白质的未来化学蛋白质组学分析以剖析心脏肿瘤学的潜在机制。
