Experimental Determination of the Förster Distance for Two Commonly Used Bioluminescent Resonance Energy Transfer Pairs

• Published in: Analytical chemistry (Washington) Vol. 82; no. 1; pp. 432 - 435
• Main Authors: DACRES, H, WANG, J, DUMANCIC, M. M, TROWELL, S. C
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01-01-2010
• Subjects: Analytical chemistry; Biological and medical sciences; Bioluminescence; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biosensors; Biotechnology; Chemistry; Electron Transport; Exact sciences and technology; Experiments; Fluorescence Resonance Energy Transfer - methods; Fundamental and applied biological sciences. Psychology; General, instrumentation; Luminescent Measurements - methods; Methods. Procedures. Technologies; Optimization; Sensitivity and Specificity; Various methods and equipments; Fluorophore; Efficiency; Resonant energy transfer; Biosensor; Monitoring; Optimization
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac9022956

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.