TY  - JOUR
A1  - Röthlein, Christoph
A1  - Miettinen, Markus S.
A1  - Ignatova, Zoya
T1  - A flexible approach to assess fluorescence decay functions in complex energy transfer systems
T2  - BMC biophysics
N2  - Background: Time-correlated Forster resonance energy transfer (FRET) probes molecular distances with greater accuracy than intensity-based calculation of FRET efficiency and provides a powerful tool to study biomolecular structure and dynamics. Moreover, time-correlated photon count measurements bear additional information on the variety of donor surroundings allowing more detailed differentiation between distinct structural geometries which are typically inaccessible to general fitting solutions.
 Results: Here we develop a new approach based on Monte Carlo simulations of time-correlated FRET events to estimate the time-correlated single photon counts (TCSPC) histograms in complex systems. This simulation solution assesses the full statistics of time-correlated photon counts and distance distributions of fluorescently labeled biomolecules. The simulations are consistent with the theoretical predictions of the dye behavior in FRET systems with defined dye distances and measurements of randomly distributed dye solutions. We validate the simulation results using a highly heterogeneous aggregation system and explore the conditions to use this tool in complex systems.
 Conclusion: This approach is powerful in distinguishing distance distributions in a wide variety of experimental setups, thus providing a versatile tool to accurately distinguish between different structural assemblies in highly complex systems.
KW  - Time resolved FRET
KW  - Monte-Carlo simulations
KW  - Complex heterogeneous systems
KW  - Protein aggregation
Y1  - 2015
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/39014
SN  - 2046-1682
VL  - 8
PB  - BioMed Central
CY  - London
ER  -