[EN] ACTIVE SITE PROBES<br/>[FR] SONDES À SITE ACTIF
申请人:ISIS INNOVATION
公开号:WO2013144656A1
公开(公告)日:2013-10-03
According to the present invention, there is provided a probe for use in the detection of a deubiquitinating enzyme, wherein the probe comprises: a protein comprising an amino acid sequence of a ubiquitin or ubiquitin-like protein; and a group of the formula -L-Z attached at the carboxy terminus of said amino acid sequence, wherein L is a linker group comprising an electrophilic moiety which is capable of covalently binding to an active site of a deubiquitinating enzyme; and Z is a detectable label or a conjugated substance; and wherein the probe comprises at least one detectable label. Also provided are processes for producing said probes, methods of using said probes, and kits comprising said probes.
Fluorescence-based active site probes for profiling deubiquitinating enzymes
作者:Joanna F. McGouran、Holger B. Kramer、Mukram M. Mackeen、Katalin di Gleria、Mikael Altun、Benedikt M. Kessler
DOI:10.1039/c2ob25258a
日期:——
Novel ubiquitin-based active site probes including a fluorescent tag have been developed and evaluated. A new, functionalizable electrophilic trap is utilized allowing for late stage diversification of the probe. Attachment of fluorescent dyes allowed direct detection of endogenous deubiquitinating enzyme (DUB) activities in cell extracts by in-gel fluorescence imaging.