Analysis of Fluorescence Lifetime and Energy Transfer Efficiency in Single-Molecule Photon Trajectories of Fast-Folding Proteins

The Journal of Physical Chemistry. B
Hoi Sung ChungIrina V Gopich

Abstract

In single-molecule Förster resonance energy transfer (FRET) spectroscopy, the dynamics of molecular processes are usually determined by analyzing the fluorescence intensity of donor and acceptor dyes. Since FRET efficiency is related to fluorescence lifetimes, additional information can be extracted by analyzing fluorescence intensity and lifetime together. For fast processes where individual states are not well separated in a trajectory, it is not easy to obtain the lifetime information. Here, we present analysis methods to utilize fluorescence lifetime information from single-molecule FRET experiments, and apply these methods to three fast-folding, two-state proteins. By constructing 2D FRET efficiency-lifetime histograms, the correlation can be visualized between the FRET efficiency and fluorescence lifetimes in the presence of the submicrosecond to millisecond dynamics. We extend the previously developed method for analyzing delay times of donor photons to include acceptor delay times. To determine the kinetics and lifetime parameters accurately, we used a maximum likelihood method. We found that acceptor blinking can lead to inaccurate parameters in the donor delay time analysis. This problem can be solved by incorporating...Continue Reading

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Citations

Feb 22, 2018·Nature·Alessandro BorgiaBenjamin Schuler
Aug 2, 2017·Proceedings of the National Academy of Sciences of the United States of America·Hoi Sung ChungJohn M Louis
Apr 2, 2018·The Journal of Chemical Physics·Wenwei ZhengRobert B Best
May 30, 2017·Journal of Molecular Biology·Hoi Sung Chung
Oct 8, 2019·Biophysical Journal·Janghyun YooHoi Sung Chung
Apr 9, 2021·Methods : a Companion to Methods in Enzymology·Jhullian J AlstonAlex S Holehouse
Sep 20, 2018·The Journal of Physical Chemistry. B·Janghyun YooHoi Sung Chung
May 17, 2018·ACS Applied Materials & Interfaces·Andres F Chaparro SosaDaniel K Schwartz
Sep 2, 2021·JACS Au·Fabian DingfelderBenjamin Schuler

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