Quantitative intensity-based FRET approaches--a comparative snapshot.

Abstract:

:Förster resonance energy transfer (FRET) has become an important tool for analyzing different aspects of interactions among biological macromolecules in their native environments. FRET analysis has also been successfully applied to study the spatiotemporal regulation of various cellular processes using genetically encoded FRET-based biosensors. A variety of procedures have been described for measuring FRET efficiency or the relative abundance of donor-acceptor complexes, based on analysis of the donor fluorescence lifetime or the spectrally resolved fluorescence intensity. The latter methods are preferable if one wants to not only quantify the apparent FRET efficiencies but also calculate donor-acceptor stoichiometry and observe fast dynamic changes in the interactions among donor and acceptor molecules in live cells. This review focuses on a comparison of the available intensity-based approaches used to measure FRET. We discuss their strengths and weaknesses in terms of FRET quantification, and provide several examples of biological applications.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Zeug A,Woehler A,Neher E,Ponimaskin EG

doi

10.1016/j.bpj.2012.09.031

subject

Has Abstract

pub_date

2012-11-07 00:00:00

pages

1821-7

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(12)01072-7

journal_volume

103

pub_type

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