Dissection of Interaction Kinetics through Single-Molecule Interaction Simulation

The ability to extract
kinetic interaction parameters from single-molecule
fluorescence resonance energy transfer trajectories without the need
for solving complex single-molecule differential equations has the
potential to address some of the critical biophysical questions. Here,
we provide a noise-free single-molecule interaction simulation (SMIS)
tool to give the expected dwell-time distributions and relative populations
of each FRET level based on the assigned kinetic model and to dissect
kinetic interaction parameters from single-molecule FRET trajectories.
The method provides the expected dwell-time distributions, average
transition rates, and relative populations of each FRET level based
on the assigned kinetic model. By comparing with ground truth data
and experimental data, we demonstrated that SMIS is useful to quantify
the interaction kinetic rate constants without using the traditional
single-molecule analytical solution approach.