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Cell-Permeable Stimuli-Responsive Ubiquitin Probe for Time- Resolved Monitoring of Substrate Ubiquitination in Live Cells

JACS Au. 2023-10; 
Lu-Jun Liang, Yu Wang, Xiao Hua, Rujing Yuan, Qiong Xia, Rongtian Wang, Chuntong Li, Guo-Chao Chu, Lei Liu, and Yi-Ming Li
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Abstract

Dynamic monitoring of intracellular ubiquitin (Ub) conjugates is instrumental to understanding the Ub regulatory machinery. Although many biochemical approaches have been developed to characterize protein ubiquitination, chemical tools capable of temporal resolution probing of ubiquitination events remain to be developed. Here, we report the development of the first cell-permeable and stimuli-responsive Ub probe and its application for the temporal resolution profiling of ubiquitinated substrates in live cells. The probe carrying the photolabile group N- (2-nitrobenzyl)-Gly (Nbg) on the amide bond between Ub Gly75 and Gly76 is readily prepared through chemical synthesis and can be delivered to live cells by c... More

Keywords

protein ubiquitination, chemical protein synthesis, Ub probes, photocaged probes, proteomic profiling