Chemical Synthesis of Activity-Based Diubiquitin Probes

Guorui Li; Libo Yuan; Zhihao Zhuang

doi:10.1007/978-1-4939-6539-7_16

Chemical Synthesis of Activity-Based Diubiquitin Probes

Methods Mol Biol. 2017:1513:223-232. doi: 10.1007/978-1-4939-6539-7_16.

Authors

Guorui Li¹, Libo Yuan¹, Zhihao Zhuang²

Affiliations

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.

PMID: 27807841
DOI: 10.1007/978-1-4939-6539-7_16

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.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Benzene Derivatives / chemistry*
Catalytic Domain
Chemistry Techniques, Synthetic*
Cysteine / chemistry
Deubiquitinating Enzymes / analysis*
Deubiquitinating Enzymes / metabolism
Humans
Mesna / chemistry
Molecular Probes / chemical synthesis*
Molecular Probes / chemistry
Plasmids / chemistry
Plasmids / metabolism
Ubiquitin / metabolism*
Ubiquitination

Substances

Benzene Derivatives
Molecular Probes
Ubiquitin
Deubiquitinating Enzymes
Cysteine
Mesna

Grants and funding

R21 NS085509/NS/NINDS NIH HHS/United States