Characterizing Single-Molecule FRET Dynamics with Probability Distribution Analysis
| Autor: | Yusdi Santoso, Achillefs N. Kapanidis, Joseph P. Torella |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Stereochemistry
Monte Carlo method 01 natural sciences Resonance (particle physics) 03 medical and health sciences Histogram 0103 physical sciences Fluorescence Resonance Energy Transfer Physical and Theoretical Chemistry 030304 developmental biology Klenow fragment Probability 0303 health sciences Quantitative Biology::Biomolecules 010304 chemical physics biology Chemistry Inverted Repeat Sequences Single-molecule FRET DNA DNA Polymerase I Atomic and Molecular Physics and Optics Förster resonance energy transfer biology.protein Probability distribution Nucleic Acid Conformation DNA polymerase I Biological system Monte Carlo Method |
| Zdroj: | ChemPhysChem ChemPhysChem; Vol 11 |
| ISSN: | 1439-7641 1439-4235 |
| DOI: | 10.1002/cphc.201000129 |
| Popis: | Probability distribution analysis (PDA) is a recently developed statistical tool for predicting the shapes of single-molecule fluorescence resonance energy transfer (smFRET) histograms, which allows the identification of single or multiple static molecular species within a single histogram. We used a generalized PDA method to predict the shapes of FRET histograms for molecules interconverting dynamically between multiple states. This method is tested on a series of model systems, including both static DNA fragments and dynamic DNA hairpins. By fitting the shape of this expected distribution to experimental data, the timescale of hairpin conformational fluctuations can be recovered, in good agreement with earlier published results obtained using different techniques. This method is also applied to studying the conformational fluctuations in the unliganded Klenow fragment (KF) of Escherichia coli DNA polymerase I, which allows both confirmation of the consistency of a simple, two-state kinetic model with the observed smFRET distribution of unliganded KF and extraction of a millisecond fluctuation timescale, in good agreement with rates reported elsewhere. We expect this method to be useful in extracting rates from processes exhibiting dynamic FRET, and in hypothesis-testing models of conformational dynamics against experimental data. |
| Databáze: | OpenAIRE |
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