Synthesis and analysis of K11-linked ubiquitin chains

Methods Mol Biol. 2012;832:219-28. doi: 10.1007/978-1-61779-474-2_15.

Abstract

Much has been learned about protein ubiquitination by studying the structural, biochemical, and biophysical properties of ubiquitin chains in vitro. However, these analyses were limited to K48-, K63-linked, and linear ubiquitin chains. Only recently, enzymatic and chemical assembly systems for the remaining chain types have been developed. Here, we describe a method to synthesise K11-linked ubiquitin chains in vitro by exploiting the intrinsic K11-specificity of the E2 enzyme UBE2S.

MeSH terms

  • Cloning, Molecular / methods
  • Humans
  • Polyubiquitin / chemical synthesis*
  • Polyubiquitin / chemistry
  • Recombinant Fusion Proteins / genetics
  • Substrate Specificity
  • Ubiquitin-Conjugating Enzymes / genetics
  • Ubiquitin-Conjugating Enzymes / metabolism*
  • Ubiquitination

Substances

  • Recombinant Fusion Proteins
  • Polyubiquitin
  • Ube2S protein, human
  • Ubiquitin-Conjugating Enzymes