Cancer Gene Networks pp 223-232 | Cite as
Chemical Synthesis of Activity-Based Diubiquitin Probes
Protocol
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Abstract
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.
Key words
Activity-based probe Diubiquitin probe Chemical ligation DUB Ubiquitin chain Linkage specificityNotes
Acknowledgment
This work was supported, in part, by the US NIH grants GM097468 and NS085509 to Z. Zhuang.
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