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Title: Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency

Abstract

Forster resonance energy transfer (FRET) is a widely used single-molecule technique for measuring nanoscale distances from changes in the non-radiative transfer of energy between donor and acceptor fluorophores. For macromolecules and complexes this observed transfer efficiency is used to infer changes in molecular conformation under differing experimental conditions. But, sometimes shifts are observed in the FRET efficiency even when there is strong experimental evidence that the molecular conformational state is unchanged. Here, we investigate ways in which such discrepancies can arise from kinetic effects. We show that significant shifts can arise from the interplay between excitation kinetics, orientation diffusion of fluorophores, separation diffusion of fluorophores, and non-emitting quenching.

Authors:
 [1]; ORCiD logo [2];  [3]
  1. Univ. of California, Santa Barbara, CA (United States). Dept. of Mathematics
  2. Univ. of California, Santa Barbara, CA (United States). Dept. of Mathematics, Dept. of Mechanical Engineering
  3. National Research Council, Rome (Italy)
Publication Date:
Research Org.:
Univ. of California, Santa Barbara, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Advanced Scientific Computing Research (ASCR) (SC-21); National Science Foundation (NSF)
OSTI Identifier:
1372594
Alternate Identifier(s):
OSTI ID: 1389573
Grant/Contract Number:
SC0009254; 0956210
Resource Type:
Journal Article: Published Article
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 12; Journal Issue: 5; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSICS; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Wallace, Bram, Atzberger, Paul J., and D’Auria, Sabato. Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency. United States: N. p., 2017. Web. doi:10.1371/journal.pone.0177122.
Wallace, Bram, Atzberger, Paul J., & D’Auria, Sabato. Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency. United States. doi:10.1371/journal.pone.0177122.
Wallace, Bram, Atzberger, Paul J., and D’Auria, Sabato. Fri . "Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency". United States. doi:10.1371/journal.pone.0177122.
@article{osti_1372594,
title = {Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency},
author = {Wallace, Bram and Atzberger, Paul J. and D’Auria, Sabato},
abstractNote = {Forster resonance energy transfer (FRET) is a widely used single-molecule technique for measuring nanoscale distances from changes in the non-radiative transfer of energy between donor and acceptor fluorophores. For macromolecules and complexes this observed transfer efficiency is used to infer changes in molecular conformation under differing experimental conditions. But, sometimes shifts are observed in the FRET efficiency even when there is strong experimental evidence that the molecular conformational state is unchanged. Here, we investigate ways in which such discrepancies can arise from kinetic effects. We show that significant shifts can arise from the interplay between excitation kinetics, orientation diffusion of fluorophores, separation diffusion of fluorophores, and non-emitting quenching.},
doi = {10.1371/journal.pone.0177122},
journal = {PLoS ONE},
number = 5,
volume = 12,
place = {United States},
year = {Fri May 19 00:00:00 EDT 2017},
month = {Fri May 19 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1371/journal.pone.0177122

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