@misc{SchulerLipmanSteinbachetal.2005,
  author    = {Schuler, Benjamin and Lipman, Everett A. and Steinbach, Peter J. and Kumke, Michael Uwe and Eaton, William A.},
  title     = {Polyproline and the "spectroscopic ruler" revisited with single-molecule fluorescence},
  url       = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-12229},
  year      = {2005},
  abstract  = {To determine whether F{\"o}rster resonance energy transfer (FRET) measurements can provide quantitative distance information in single-molecule fluorescence experiments on polypeptides, we measured FRET efficiency distributions for donor and acceptor dyes attached to the ends of freely diffusing polyproline molecules of various lengths. The observed mean FRET efficiencies agree with those determined from ensemble lifetime measurements but differ considerably from the values expected from F{\"o}rster theory, with polyproline treated as a rigid rod. At donor-acceptor distances much less than the F{\"o}rster radius R0, the observed efficiencies are lower than predicted, whereas at distances comparable to and greater than R0, they are much higher. Two possible contributions to the former are incomplete orientational averaging during the donor lifetime and, because of the large size of the dyes, breakdown of the point-dipole approximation assumed in F{\"o}rster theory. End-to-end distance distributions and correlation times obtained from Langevin molecular dynamics simulations suggest that the differences for the longer polyproline peptides can be explained by chain bending, which considerably shortens the donor-acceptor distances.},
  language  = {en}
}