A bifunctional molecule-assisted synthesis of mimics for use in probing the ubiquitination system
Qingyun Zheng, Tian Wang, Junxiong Mao, Guochao Chu, Lujun Liang, Yangwode Jing, Chong Zuo, Yuanyuan Yu, Honggang Hu, Man Pan
Published: 2022-11-01 DOI: 10.1038/s41596-022-00761-z





























Extended
Extended Data Fig. 1 Intermediate products of the Acm-deprotection reaction and CAET-assisted NCL reaction for preparation of Ubc2-Ub- K48 Ub-Y/degron.
a , RP-HPLC trace (214 nm) and ESI-MS of purified product 6. b , RP-HPLC trace (214 nm) and ESI-MS of purified product 8 .
Extended Data Fig. 2 LC–MS characterization of Ub-K27C-CAET module and monitoring the reaction efficiency of the DUB intermediate mimic.
LC–MS characterization of Ub-K27C-CAET module and monitoring the reaction efficiency of the DUB intermediate mimic. a , RP-HPLC trace (214 nm) and ESI-MS of purified product 12. b , SDS–PAGE analysis of the activity-based cross-linking between the enzyme Otud2 and K27-CAET-AT2 diUb.