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Reliable State Identification and State Transition Detection in Fluorescence Intensity-Based Single-Molecule Förster Resonance Energy-Transfer Data

Hadzic, Mélodie C A S; Börner, Richard; König, Sebastian L B; Kowerko, Danny; Sigel, Roland K O (2018). Reliable State Identification and State Transition Detection in Fluorescence Intensity-Based Single-Molecule Förster Resonance Energy-Transfer Data. Journal of Physical Chemistry B, 122(23):6134-6147.

Abstract

Single-molecule Förster resonance energy transfer (smFRET) is a powerful technique to probe biomolecular structure and dynamics. A popular implementation of smFRET consists of recording fluorescence intensity time traces of surface-immobilized, chromophore-tagged molecules. This approach generates large and complex data sets, the analysis of which is to date not standardized. Here, we address a key challenge in smFRET data analysis: the generation of thermodynamic and kinetic models that describe with statistical rigor the behavior of FRET trajectories recorded from surface-tethered biomolecules in terms of the number of FRET states, the corresponding mean FRET values, and the kinetic rates at which they interconvert. For this purpose, we first perform Monte Carlo simulations to generate smFRET trajectories, in which a relevant space of experimental parameters is explored. Then, we provide an account on current strategies to achieve such model selection, as well as a quantitative assessment of their performances. Specifically, we evaluate the performance of each algorithm (change-point analysis, STaSI, HaMMy, vbFRET, and ebFRET) with respect to accuracy, reproducibility, and computing time, which yields a range of algorithm-specific referential benchmarks for various data qualities. Data simulation and analysis were performed with our MATLAB-based multifunctional analysis software for handling smFRET data (MASH-FRET).

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Scopus Subject Areas:Physical Sciences > Physical and Theoretical Chemistry
Physical Sciences > Surfaces, Coatings and Films
Physical Sciences > Materials Chemistry
Uncontrolled Keywords:Physical and Theoretical Chemistry, Materials Chemistry, Surfaces, Coatings and Films
Language:English
Date:2018
Deposited On:12 Jun 2018 15:55
Last Modified:18 Jan 2025 02:40
Publisher:American Chemical Society (ACS)
ISSN:1520-5207
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1021/acs.jpcb.7b12483
PubMed ID:29737844
Project Information:
  • Funder: FP7
  • Grant ID: 259092
  • Project Title: MIRNA - Metal Ions and Metal Ion Complexes Guiding Folding and Function of Single RNA Molecules
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