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Recently developed statistical methods by Gopich and Szabo were used to extract folding and unfolding rate coefficients from single-molecule Förster resonance energy transfer (FRET) data for proteins with kinetics too fast to measure waiting time distributions. Two types of experiments and two different analyses were performed. In one experiment bursts of photons were collected from donor and acceptor fluorophores attached to a 73-residue protein, α3D, freely diffusing through the illuminated volume of a confocal microscope system. In the second, the protein was immobilized by linkage to a surface, and photons were collected until one of the fluorophores bleached. Folding and unfolding rate coefficients and mean FRET efficiencies for the folded and unfolded subpopulations were obtained from a photon by photon analysis of the trajectories using a maximum likelihood method. The ability of the method to describe the data in terms of a two-state model was checked by recoloring the photon trajectories with th...
| Authors: | Hoi Sung Chung; Irina V. Gopich; Kevin McHale; Troy Cellmer; John M. Louis; William A. Eaton | |
| Journal: | Journal of Physical Chemistry A | |
| Year: | 2010 | |
| DOI: | 10.1021/jp1009669 | |
| Publication date: | 28-05-2010 |
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