Activation of procaspases by FK506 binding protein-mediated oligomerization

Sci STKE. 2003 Jan 28;2003(167):PL1. doi: 10.1126/stke.2003.167.pl1.

Abstract

Oligomerization is an important biological mechanism for regulating signal transduction. Activation of caspases during apoptosis is triggered by adaptor protein-mediated oligomerization of initiator procaspases. To facilitate the study of initiator caspase activation, a system that allows inducible activation of various caspases both in vitro and in vivo is highly desired. Here we describe such a caspase activation system that is based on FK506 binding protein (FKBP)-mediated oligomerization. The NH(2)-terminal prodomains of initiator procaspases that facilitate the interaction between procaspases and their adaptor proteins are replaced by a derivative of FKBP called Fv. The Fv-caspase fusions can then be dimerized by a synthetic divalent Fv ligand, AP20187, which binds strongly to Fv but weakly to the endogenous FKBPs. This FKBP-based system may be widely applicable to the study of the regulation and functions of caspases.

MeSH terms

  • Caspase 8
  • Caspase 9
  • Caspases / biosynthesis
  • Caspases / genetics
  • Caspases / metabolism*
  • Cell Line
  • DNA, Complementary / genetics
  • Dimerization
  • Enzyme Activation
  • Enzyme Precursors / metabolism*
  • Gene Expression Regulation, Enzymologic / genetics
  • Genetic Vectors / genetics
  • HeLa Cells
  • Humans
  • Kidney
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Tacrolimus / analogs & derivatives*
  • Tacrolimus / metabolism
  • Tacrolimus Binding Proteins / physiology*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • AP20187
  • DNA, Complementary
  • Enzyme Precursors
  • Recombinant Fusion Proteins
  • CASP8 protein, human
  • CASP9 protein, human
  • Caspase 8
  • Caspase 9
  • Caspases
  • Tacrolimus Binding Proteins
  • Tacrolimus