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Abstract
This chapter discusses the applications of bioluminescence resonance energy transfer (BRET) in biological problems. BRET has potential advantages over Förster resonance energy transfer (FRET) because it does not require the use of excitation illumination; BRET is superior for cells that are either photoresponsive (for example, retina or any photoreceptive tissue) or damaged by the wavelength of light used to excite FRET. Photobleaching of the fluorophores can be a serious limitation of FRET, but it is irrelevant to BRET. Cells that have significant autofluorescence are better assayed by BRET than by FRET. This is particularly true for highly autofluorescent tissue, but all cells are autofluorescent to a degree because of ubiquitous fluorescent molecules such as NADH, collagen, and flavins. In plant cells, BRET is particularly promising because the highly fluorescent photosynthetic pigments and cell-wall compounds prevalent in plants can interfere with FRET-based assays. The first application of BRET to plant cells has demonstrated the interaction of the phototransduction molecule COPl in onion and Arabidopsis.
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