UBP43, an ISG15-specific deconjugating enzyme: expression, purification, and enzymatic assays

Methods Enzymol. 2005:398:491-9. doi: 10.1016/S0076-6879(05)98040-3.

Abstract

UBP43 is the only deconjugating enzyme for the protein ISGylation system thus far identified. UBP43 activity is not critical for precursor processing, as UBP43-deficent cells can generate ISG15 conjugates upon type I interferon treatment. However, UBP43 deficiency caused a defect in the negative regulation of type I interferon signaling, resulting in enhanced and prolonged activation of Jak/Stat upon type I interferon treatment. This chapter describes the expression, purification, and enzymatic assays for UBP43.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cloning, Molecular
  • Cytokines / genetics
  • Cytokines / metabolism*
  • Endopeptidases / biosynthesis*
  • Endopeptidases / genetics
  • Endopeptidases / isolation & purification*
  • Endopeptidases / metabolism
  • Humans
  • Mice
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Ubiquitin Thiolesterase
  • Ubiquitins / genetics
  • Ubiquitins / metabolism
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Cytokines
  • Recombinant Fusion Proteins
  • Ubiquitins
  • ISG15 protein, human
  • beta-Galactosidase
  • Endopeptidases
  • Usp18 protein, mouse
  • USP18 protein, human
  • Ubiquitin Thiolesterase