Nature Biotechnology
16, 547 - 552 (1998)
doi:10.1038/nbt0698-547
Bcl-2 and Bax interactions in mitochondria probed with green fluorescent protein and fluorescence resonance energy transferNupam P. Mahajan1, Katrina Linder1,4, Gail Berry1, Gerald W. Gordon1, Roger Heim3
& Brian Herman1,2,*
1Department of Cell Biology and Anatomy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
2The Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
3Aurora Bioscience, La Jolla, CA 92093.
4Current address: Glaxo Wellcome Inc., Research Triangle Park, NC 27710.
*Corresponding author (e-mail: hertnanb@UTHSCSA.EDU). 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.
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