Efficient use of single molecule time traces to resolve kinetic rates, models and uncertainties
arXiv:1709.08270 · doi:10.1063/1.5006604
Abstract
Single molecule time traces reveal the time evolution of unsynchronized kinetic systems. Especially single molecule Förster resonance energy transfer (smFRET) provides access to enzymatically important timescales, combined with molecular distance resolution and minimal interference with the sample. Yet the kinetic analysis of smFRET time traces is complicated by experimental shortcomings - such as photo-bleaching and noise. Here we recapitulate the fundamental limits of single molecule fluorescence that render the classic, dwell-time based kinetic analysis unsuitable. In contrast, our Single Molecule Analysis of Complex Kinetic Sequences (SMACKS) considers every data point and combines the information of many short traces in one global kinetic rate model. We demonstrate the potential of SMACKS by resolving the small kinetic effects caused by different ionic strengths in the chaperone protein Hsp90. These results show an unexpected interrelation between conformational dynamics and ATPase activity in Hsp90.
arXiv comment17 pages, 6 figures
References in corpus (4)
- What can one learn from two-state single molecule trajectories?
- Experiment-friendly kinetic analysis of single molecule data in and out of equilibrium
- Inferring hidden Markov models from noisy time sequences: a method to alleviate degeneracy in molecular dynamics
- Can one identify non-equilibrium in a three-state system by analyzing two-state trajectories?