我们提出了天然化学连接柄的开发,该柄还用作掩蔽的亲电子试剂,在需要时可以在合成过程中释放。该柄因此可用于合成复杂的基于活性的探针。我们描述了该手柄在生成基于连锁特异性活性的去泛素化酶探针中的用途,该探针包含底物上下文并紧密模拟天然泛素异肽键。我们已经基于所有七个异肽连接的二泛素拓扑异构体产生了基于活性的探针,并证明了它们的结构完整性和以连接特异性方式标记DUB的能力。
We present the development of a native chemical ligation handle that also functions as a masked electrophile that can be liberated during synthesis when required. This handle can thus be used for the synthesis of complex activity-based probes. We describe the use of this handle in the generation of linkage-specific activity-based deubiquitylating enzyme probes that contain substrate context and closely mimic the native ubiquitin isopeptide linkage. We have generated activity-based probes based on all seven isopeptide-linked diubiquitin topoisomers and demonstrated their structural integrity and ability to label DUBs in a linkage-specific manner.