A simple "proximity" correction for Förster resonance energy transfer efficiency determination in membranes using lifetime measurements

Yevgen O. Posokhov, Mikhail Merzlyakov, Kalina A Hristova, Alexey S. Ladokhin

Research output: Contribution to journalArticle

Abstract

Accurate measurements of oligomerization in membranes by Förster resonance energy transfer (FRET) are always compromised by a substantial contribution from random chance colocalization of donors and acceptors. Recently, Li and coworkers demonstrated the use of computer simulation in estimating the contribution of this "proximity" component to correct the FRET efficiency and estimate the free energy of dimer formation of the G380R mutants of fibroblast growth factor receptor 3 (FGFR3) transmembrane domain immersed into lipid bilayer. Because tight dimerization will result in complete energy transfer from donor to acceptor, we have used the same experimental system of fluorescein- and rhodamine-labeled G380R mutants of FGFR3 for the experimental assessment of the proximity FRET corrections using fluorescence lifetime measurements. The experimental proximity FRET correction, based on time-resolved fluorescence measurements, is expected to have general advantages over theoretical correction, especially in the case of nonrandomly distributed monomers.

Original languageEnglish (US)
Pages (from-to)134-136
Number of pages3
JournalAnalytical Biochemistry
Volume380
Issue number1
DOIs
StatePublished - Sep 1 2008

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Energy Transfer
Energy transfer
Membranes
Receptor, Fibroblast Growth Factor, Type 3
Fluorescence
Oligomerization
Lipid bilayers
Rhodamines
Dimerization
Lipid Bilayers
Fluorescein
Dimers
Computer Simulation
Free energy
Monomers
Computer simulation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A simple "proximity" correction for Förster resonance energy transfer efficiency determination in membranes using lifetime measurements. / Posokhov, Yevgen O.; Merzlyakov, Mikhail; Hristova, Kalina A; Ladokhin, Alexey S.

In: Analytical Biochemistry, Vol. 380, No. 1, 01.09.2008, p. 134-136.

Research output: Contribution to journalArticle

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