Activity-based diubiquitin probes are highly useful in probing the deubiquitinase (DUB) activity and ubiquitin chain linkage specificity. Here we describe a detailed protocol to synthesize a new class of diubiquitin DUB probes. In this method, two ubiquitin moieties are connected through a linker resembling the native linkage in size and containing a Michael acceptor for trapping the DUB active-site cysteine. Detailed procedures for generating the linker molecule are also described.
Li, G., Yuan, L., & Zhuang, Z. (2017). Chemical synthesis of activity-based Diubiquitin probes. In Methods in Molecular Biology (Vol. 1513, pp. 223–232). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6539-7_16