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A Native Chemical Ligation Handle that Enables the Synthesis of Advanced Activity‐Based Probes: Diubiquitin as a Case Study
Author(s) -
Mulder Monique P. C.,
El Oualid Farid,
ter Beek Jarno,
Ovaa Huib
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402012
Subject(s) - native chemical ligation , electrophile , context (archaeology) , chemistry , chemical ligation , ubiquitin , chemical synthesis , substrate (aquarium) , combinatorial chemistry , linkage (software) , biochemistry , biology , catalysis , gene , in vitro , paleontology , ecology , peptide
We present the development of a native chemical ligation handle that also functions as a masked electrophile that can be liberated during synthesis when required. This handle can thus be used for the synthesis of complex activity‐based probes. We describe the use of this handle in the generation of linkage‐specific activity‐based deubiquitylating enzyme probes that contain substrate context and closely mimic the native ubiquitin isopeptide linkage. We have generated activity‐based probes based on all seven isopeptide‐linked diubiquitin topoisomers and demonstrated their structural integrity and ability to label DUBs in a linkage‐specific manner.