Expression and purification of human interleukin-1beta converting enzyme from Trichoplusia ni insect cells using a baculovirus expression system

Protein Expr Purif. 1997 Feb;9(1):69-75. doi: 10.1006/prep.1996.0686.

Abstract

Employing a baculovirus expression system, human interleukin-1beta converting enzyme (ICE) has been expressed in Trichoplusia ni High-Five insect cells and purified. ICE was expressed with an N-terminal T7 epitope, thus allowing its purification using immobilized anti-T7 antibodies. The recombinant ICE was purified to >95% homogeneity, the one minor contaminant being the baculovirus anti-apoptotic protein (P35) which was copurified. The purified recombinant ICE was biologically active, cleaving both pIL-1beta and poly(ADP-ribose) polymerase substrates. The kinetic properties of the purified recombinant ICE compare favorably with native ICE purified from a THP-1 monocytic line.

MeSH terms

  • Animals
  • Bacteriophage T7 / genetics
  • Bacteriophage T7 / immunology
  • Caspase 1
  • Cysteine Endopeptidases / biosynthesis*
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / isolation & purification
  • Epitopes
  • Gene Expression
  • Genetic Vectors
  • Humans
  • Inhibitor of Apoptosis Proteins
  • Kinetics
  • Moths / cytology
  • Moths / virology
  • Nucleopolyhedroviruses / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Sequence Analysis
  • Viral Proteins / isolation & purification

Substances

  • Epitopes
  • Inhibitor of Apoptosis Proteins
  • Recombinant Fusion Proteins
  • Viral Proteins
  • inhibitor of apoptosis, Nucleopolyhedrovirus
  • Cysteine Endopeptidases
  • Caspase 1