Conversion of Bfl-1, an anti-apoptotic Bcl-2 family protein, to a potent pro-apoptotic protein by fusion with green fluorescent protein (GFP)

FEBS Lett. 2003 Sep 11;551(1-3):29-36. doi: 10.1016/s0014-5793(03)00872-x.

Abstract

Human Bfl-1 is an anti-apoptotic Bcl-2 family member. Here, we found that Bfl-1 was converted into a potent death-promoting protein by green fluorescent protein (GFP) fusion with its N-terminus. The transient expression of GFP-Bfl-1 induced cytochrome c release and triggered apoptosis in 293T cells, which depended on the mitochondrial localization of GFP-Bfl-1. Apoptosis induced by GFP-Bfl-1 was significantly blocked by the pan-caspase inhibitor carbobenzoxy-Val-Ala-Asp-fluoromethyl ketone, but was not blocked by either Bcl-xL or Bfl-1. Our findings provide a useful model for understanding the structural basis of Bcl-2 family proteins that act in an opposite way despite sharing structural similarity between anti-apoptotic and pro-apoptotic proteins.

MeSH terms

  • Apoptosis*
  • Caspases / physiology
  • Cell Line
  • Cytochrome c Group / metabolism
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins / genetics
  • Minor Histocompatibility Antigens
  • Mitochondria / chemistry
  • Mitochondria / metabolism
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-bcl-2 / chemistry*
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / antagonists & inhibitors
  • Recombinant Fusion Proteins / metabolism
  • bcl-X Protein

Substances

  • BCL2-related protein A1
  • BCL2L1 protein, human
  • Cytochrome c Group
  • Luminescent Proteins
  • Minor Histocompatibility Antigens
  • Proto-Oncogene Proteins c-bcl-2
  • Recombinant Fusion Proteins
  • bcl-X Protein
  • Green Fluorescent Proteins
  • Caspases