Table 2. Measures of Fluorescence Lifetime of P450:eGFP and FRET Evaluation of P450:eGFP/P450:mRFP1 Associations.
| Expressed Proteins |
Fluorescence Lifetime (ns) |
FRET Efficiency |
||||
|---|---|---|---|---|---|---|
| Donor | Acceptor | n | Average | sd | % | sd |
| CYP73A5:eGFP | / | 121 | 2.05 | 0.09 | – | – |
| mRFP1 | 30 | 1.97 | 0.08 | 3.6 | 0.2 | |
| Anchored-mRFP1 | 20 | 1.99 | 0.31 | 2.7 | 0.4 | |
| CYP71B31:mRFP1 | 17 | 1.96 | 0.07 | 4.5 | 0.3 | |
| CYP98A3:mRFP1 | 22 | 1.70 | 0.17 | 17.2 | 1.8 | |
| CYP73A5:mRFP1 | 26 | 1.86 | 0.07 | 9.3 | 0.5 | |
| CYP98A3:eGFP | / | 112 | 2.12 | 0.15 | – | – |
| mRFP1 | 17 | 2.00 | 0.14 | 5.5 | 0.5 | |
| Anchored-mRFP1 | 18 | 2.13 | 0.07 | 0.0 | 0.0 | |
| CYP71B31:mRFP1 | 8 | 2.13 | 0.23 | 0.0 | 0.1 | |
| CYP98A3:mRFP1 | 20 | 1.65 | 0.09 | 22.2 | 2.0 | |
| CYP73A5:mRFP1 | 26 | 1.92 | 0.12 | 9.1 | 0.9 | |
| CYP71B31:eGFP | / | 11 | 2.33 | 0.16 | – | – |
| CYP98A3:mRFP1 | 10 | 2.22 | 0.13 | 4.4 | 0.4 | |
| CYP73A5:mRFP1 | 8 | 2.18 | 0.12 | 6.4 | 0.6 | |
Values of P450:eGFP fluorescence lifetimes were obtained by FLIM. FRET efficiency is calculated by comparison with eGFP fluorescence lifetime measured in a control experiment in absence of the mRFP1 acceptor protein. n, number of independent cells analyzed. Slash indicates no acceptor; dash indicates no value; bold value indicates value higher than controls.