Dissecting ubiquitin signaling with linkage-defined and protease resistant ubiquitin chains

Angew Chem Int Ed Engl. 2014 Nov 17;53(47):12925-9. doi: 10.1002/anie.201407192. Epub 2014 Sep 4.

Abstract

Ubiquitylation is a complex posttranslational protein modification and deregulation of this pathway has been associated with different human disorders. Ubiquitylation comes in different flavors: Besides mono-ubiquitylation, ubiquitin chains of various topologies are formed on substrate proteins. The fate of ubiquitylated proteins is determined by the linkage-type of the attached ubiquitin chains, however, the underlying mechanism is poorly characterized. Herein, we describe a new method based on codon expansion and click-chemistry-based polymerization to generate linkage-defined ubiquitin chains that are resistant to ubiquitin-specific proteases and adopt native-like functions. The potential of these artificial chains for analyzing ubiquitin signaling is demonstrated by linkage-specific effects on cell-cycle progression.

Keywords: click chemistry; codon expansion; posttranslational modification; ubiquitin chains; unnatural amino acids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Click Chemistry
  • Escherichia coli / cytology
  • Escherichia coli / metabolism
  • Peptide Hydrolases / metabolism*
  • Polymerization
  • Signal Transduction*
  • Ubiquitin / chemistry*
  • Ubiquitin / metabolism*
  • Ubiquitination
  • Xenopus

Substances

  • Ubiquitin
  • Peptide Hydrolases