PT  - JOURNAL ARTICLE
AU  - Harris, Paul David
AU  - Lerner, Eitan
TI  - Identification and Quantification of Within-Burst Dynamics in Singly-Labeled Single-Molecule Fluorescence Lifetime Experiments
AID  - 10.1101/2022.05.23.493026
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.05.23.493026
4099  - http://biorxiv.org/content/early/2022/05/24/2022.05.23.493026.short
4100  - http://biorxiv.org/content/early/2022/05/24/2022.05.23.493026.full
AB  - Single-molecule spectroscopy has revolutionized molecular biophysics and provided means to probe how structural moieties within biomolecules spatially reorganize at different timescales. There are several single-molecule methodologies that probe local structural dynamics in the vicinity of a single dye-labeled residue, which rely on fluorescence lifetimes as readout. Nevertheless, an analytical framework to quantify dynamics in such single-molecule single-dye fluorescence bursts, at timescales of microseconds to milliseconds, has not yet been demonstrated. Here, we suggest an analytical framework for identifying and quantifying within-burst lifetime-based dynamics, such as conformational dynamics recorded in single-molecule photo-isomerization related fluorescence enhancement. After testing the capabilities of the analysis on simulations, we proceed to exhibit within-burst millisecond local structural dynamics in the unbound α-synuclein monomer. The analytical framework provided in this work paves the way for extracting a full picture of the energy landscape for the coordinate probed by fluorescence-lifetime based single-molecule measurements.Competing Interest StatementThe authors have declared no competing interest.