Expression and functional analysis of an inhibitor of apoptosis protein from Trichoplusia ni

Biochem Biophys Res Commun. 2002 May 3;293(2):675-9. doi: 10.1016/S0006-291X(02)00279-6.

Abstract

An inhibitor of the apoptosis protein (IAP) family gene from Trichoplusia ni, Tn-IAP1v, a variant of lepidopteran Tn-IAP1, was cloned by RT-PCR. There are six single nucleotide polymorphisms between the two Tn-IAP1 variants, resulting in three predicted single amino acid polymorphisms. With the GST fusion expression system, soluble recombinant Tn-IAP1v was highly expressed in Escherichia coli and then purified by affinity chromatography. Caspase inhibition assays indicated that recombinant Tn-IAP1v could specifically inhibit human caspase-9 in vitro instead of caspase-3, -7, and -8, which was further confirmed by the observation that recombinant Tn-IAP1v can directly bind caspase-9 in the protein pull-down assay. These results suggested that Tn-IAP1v might serve as an initiator caspase inhibitor in vivo in the conserved mitochondria apoptotic pathway.

MeSH terms

  • Animals
  • Caspase 9
  • Caspases / metabolism
  • Cell Line
  • Cloning, Molecular
  • Cysteine Proteinase Inhibitors / genetics
  • Cysteine Proteinase Inhibitors / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology*
  • Cytochrome c Group / antagonists & inhibitors
  • Deoxyadenine Nucleotides / antagonists & inhibitors
  • Humans
  • Inhibitor of Apoptosis Proteins
  • Insect Proteins / genetics
  • Insect Proteins / metabolism*
  • Insect Proteins / pharmacology*
  • Lepidoptera*
  • Polymorphism, Single Nucleotide
  • Proteins*
  • Ubiquitin-Protein Ligases

Substances

  • Cysteine Proteinase Inhibitors
  • Cytochrome c Group
  • Deoxyadenine Nucleotides
  • Inhibitor of Apoptosis Proteins
  • Insect Proteins
  • Proteins
  • BIRC2 protein, human
  • Ubiquitin-Protein Ligases
  • CASP9 protein, human
  • Caspase 9
  • Caspases
  • 2'-deoxyadenosine triphosphate