Ubiquitin 7-amino-4-carbamoylmethylcoumarin as an improved fluorogenic substrate for deubiquitinating enzymes

Li, Yi-Tong; Huang, Yi-Chao; Xu, Yang; Holdsworth, May; Li, Yi-Ming

Ubiquitin 7-amino-4-carbamoylmethylcoumarin as an improved fluorogenic substrate for deubiquitinating enzymes

2016

Li, Yi-Tong | Huang, Yi-Chao | Xu, Yang | Holdsworth, May | Li, Yi-Ming

A new fluorogenic substrate Ub-ACC (ubiquitin C-terminal 7-amino-4-carbamoylmethyl-coumarin) was developed for DUB (deubiquitinating enzyme) activity assays. This substrate can be synthesized with higher efficiency than the classical DUB substrate, Ub-AMC (ubiquitin C-terminal 7-amino-4-methylcoumarin). DUB assays using UCH-L3, OTUD2 and USP30 demonstrated that Ub-ACC shows nearly 2-fold higher sensitivity than Ub-AMC.

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