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Efficient Semi‐Synthesis of Atypical Ubiquitin Chains and Ubiquitin‐Based Probes Forged by Thioether Isopeptide Bonds
- Source :
- Chemistry – A European Journal. 25:16668-16675
- Publication Year :
- 2019
- Publisher :
- Wiley, 2019.
-
Abstract
- The development of powerful and general methods to acquire ubiquitin (Ub) chains has prompted the deciphering of Ub-mediated processes. Herein, the cysteine-aminoethylation assisted chemical ubiquitination (CAACU) strategy is extended and improved to enable the efficient semi-synthesis of atypical Ub chain analogues and Ub-based probes. Combining the Cys aminoethylation and the auxiliary-mediated protein ligation, several linkage- and length-defined atypical Ub chains including di-Ubs, K27C-linked tri-Ub, K11/K48C-branched tri-Ub, and even the SUMOlated Ub are successfully prepared from recombinantly expressed starting materials at about a 9-20 mg L-1 expression level. In addition, the utility of this strategy is demonstrated with the synthesis of a novel non-hydrolyzable di-Ub PA probe, which may provide a new useful tool for the mechanistic studies of deubiquitinase (DUB) recognition.
Details
- ISSN :
- 15213765 and 09476539
- Volume :
- 25
- Database :
- OpenAIRE
- Journal :
- Chemistry – A European Journal
- Accession number :
- edsair.doi.dedup.....9d1327f9c979208999dffda7a15b04bc