Abstract
Atypical ubiquitin (Ub) chains were reported to be involved in certain significant cellular processes. The development of effective methods to acquire atypical Ub chains would facilitate the deciphering of Ub-mediated processes. Herein, we reported a new enzymatic method combined with cysteine-aminoethylation assisted chemical ubiquitination (CAACU) strategy to efficiently synthesize K27-linked-mixed-triubiquitin. Compared to traditional CAACU strategy, one advantage of this method is that only one auxiliary group needs to be removed to increase the yield. By using this strategy, several mixed-triUb were easily semi-synthesized with defined-linkage.
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