Bcl-2 and bax interactions in mitochondria probed with green fluorescent protein and fluorescence resonance energy transfer

Nupam P. Mahajan, Katrina Linder, Gail Berry, Gerald W. Gordon, Roger Heim, Brian Herman

Research output: Contribution to journalArticlepeer-review

252 Scopus citations

Abstract

It has been hypothesized that interaction of Bcl-2 and Bax may regulate apoptosis. The spatial and temporal interaction of Bcl-2 and Bax at the single cell level has not, however, been demonstrated. To achieve this goal, we have developed two-fusion FRET (fluorescence resonance energy transfer). Using green fluorescent protein (GFP)-Bax and blue fluorescent protein (BFP)- Bcl-2 fusion proteins coexpressed in the same cell, we demonstrate a direct interaction between Bcl-2 and Bax in individual mitochondria. Mitochondrially localized cytochrome c-GFP and BFP-Bcl-2 showed little or no FRET, while nuclear-localized GFP-human papillomavirus E6 and BFP-Bcl-2 did not interact when coexpressed in the same cell. These findings indicate that two-fusion FRET provides an opportunity to examine the interaction between two different proteins coexpressed in single intact mammalian cells.

Original languageEnglish (US)
Pages (from-to)547-552
Number of pages6
JournalNature biotechnology
Volume16
Issue number6
DOIs
StatePublished - Jun 1 1998
Externally publishedYes

Keywords

  • Apoptosis
  • Microscopy
  • Protein interactions

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