Simultaneous Detection of Distinct Ubiquitin Chain Topologies by 19F NMR

摘要: The dynamic interplay between ubiquitin (Ub) chain construction and destruction is critical for the regulation of many cellular pathways. To understand these processes, it would be ideal to simultaneously detect different Ub chains as they are created destroyed in cell. This objective cannot achieved with existing detection strategies. Here, we report on use 19F Nuclear Magnetic Resonance (NMR) spectroscopy characterize conformationally distinct oligomers. By exploiting environmental sensitivity nucleus conformational diversity found among linkage types, can resolve NMR signals mono-Ub three di-Ub oligomers (K6, K48, K63) heterogeneous mixtures. utility this approach demonstrated by ability interrogate selectivity deubiquitinases multiple substrates real time. We also demonstrate that used discern linkages formed...