Experimental determination of the Forster distance for two commonly used bioluminescent resonance energy transfer pairs

H Dacres, J Wang, MM Dumancic… - Analytical chemistry, 2010 - ACS Publications
H Dacres, J Wang, MM Dumancic, SC Trowell
Analytical chemistry, 2010ACS Publications
Förster resonance energy transfer (RET) is the nonradiative transfer of energy from a
donor to an acceptor fluorophore. The Förster distance (R 0), being the fluorophore
separation corresponding to 50% of the maximum RET efficiency (E RET), is a critical
parameter for optimization of RET biosensors. Sensitive RET-based monitoring of molecular
rearrangements requires that the separation of the donor and acceptor RET pair is matched
to their Förster distance. Here, for the first time, we experimentally determine the Förster …
Förster resonance energy transfer (RET) is the nonradiative transfer of energy from a donor to an acceptor fluorophore. The Förster distance (R0), being the fluorophore separation corresponding to 50% of the maximum RET efficiency (ERET), is a critical parameter for optimization of RET biosensors. Sensitive RET-based monitoring of molecular rearrangements requires that the separation of the donor and acceptor RET pair is matched to their Förster distance. Here, for the first time, we experimentally determine the Förster distance for BRET1, R0 = 4.4 nm, and for BRET2, R0 = 7.5 nm. The latter is the largest reported value for a genetically encoded RET pair.
ACS Publications