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

H Dacres; J Wang; M M Dumancic; S C Trowell

doi:10.1021/ac9022956

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

Anal Chem. 2010 Jan 1;82(1):432-5. doi: 10.1021/ac9022956.

Authors

H Dacres¹, J Wang, M M Dumancic, S C Trowell

Affiliation

¹ CSIRO Food Futures Flagship & Division of Entomology, GPO Box 1700, Canberra, Australia, ACT 2601. helen.dacres@csiro.au

PMID: 19957970
DOI: 10.1021/ac9022956

Abstract

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 distance for BRET(1), R(0) = 4.4 nm, and for BRET(2), R(0) = 7.5 nm. The latter is the largest reported value for a genetically encoded RET pair.

MeSH terms

Biosensing Techniques / instrumentation*
Biosensing Techniques / methods*
Electron Transport
Fluorescence Resonance Energy Transfer / methods*
Luminescent Measurements / methods*
Sensitivity and Specificity