Chemical Synthesis of Activity-Based Diubiquitin Probes.
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.
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE, 19716, USA.
Guorui
G
Li
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE, 19716, USA.
Libo
L
Yuan
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE, 19716, USA. zzhuang@udel.edu.
Zhihao
Z
Zhuang
D001555:Benzene Derivatives / Q000737:chemistry*; D020134:Catalytic Domain; D060326:Chemistry Techniques, Synthetic; D003545:Cysteine / Q000737:chemistry; D000072017:Deubiquitinating Enzymes / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D015080:Mesna / Q000737:chemistry; D015335:Molecular Probes / Q000138:chemical synthesis* / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination
D001555:Benzene Derivatives; D015335:Molecular Probes; D025801:Ubiquitin; D000072017:Deubiquitinating Enzymes; D003545:Cysteine; D015080:Mesna
Activity-based probe; Chemical ligation; DUB; Diubiquitin probe; Linkage specificity; Ubiquitin chain