Activity-based diubiquitin probes for elucidating the linkage specificity of deubiquitinating enzymes.
We report a new class of deubiquitinating enzyme (DUB) probes that resemble the native diubiquitin with a same linkage size and contain a Michael addition acceptor for trapping the DUB active-site cysteine. Both K63- and K48-linked diubiquitin probes were generated using a facile chemical ligation method. The diUb probes were demonstrated to label DUBs from different families and revealed intrinsic linkage specificities of DUBs.
Department of Chemistry and Biochemistry, University of Delaware, 214A Drake Hall, Newark, DE 19716, USA. zzhuang@udel.edu.
Guorui
G
Li
Qin
Q
Liang
Ping
P
Gong
Adam H
AH
Tencer
Zhihao
Z
Zhuang
D000818:Animals; D020134:Catalytic Domain; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008958:Models, Molecular; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000737:chemistry*; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry*
D015335:Molecular Probes; D064570:Ubiquitin-Specific Proteases