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Methods Mol Biol. 2017;1513:223-232.

Chemical Synthesis of Activity-Based Diubiquitin Probes.

Author information

1
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE, 19716, USA.
2
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE, 19716, USA. zzhuang@udel.edu.

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.

KEYWORDS:

Activity-based probe; Chemical ligation; DUB; Diubiquitin probe; Linkage specificity; Ubiquitin chain

PMID:
27807841
DOI:
10.1007/978-1-4939-6539-7_16
[Indexed for MEDLINE]

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