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Int. J. Mol. Sci. 2012, 13(12), 16141-16156; doi:10.3390/ijms131216141

Förster Resonance Energy Transfer (FRET) between Heterogeneously Distributed Probes: Application to Lipid Nanodomains and Pores

1 Department of Biophysical Chemistry, Jaroslav Heyrovský Institute of Physical Chemistry, Academy of Sciences of the Czech Republic, Dolejškova 2155/3, Prague 8, 182 23, Czech Republic 2 Biophysical Chemistry, Department of Chemistry, Umeå University, S-901 87 Umeå, Sweden
* Author to whom correspondence should be addressed.
Received: 9 October 2012 / Revised: 14 November 2012 / Accepted: 16 November 2012 / Published: 30 November 2012
(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))
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The formation of membrane heterogeneities, e.g., lipid domains and pores, leads to a redistribution of donor (D) and acceptor (A) molecules according to their affinity to the structures formed and the remaining bilayer. If such changes sufficiently influence the Förster resonance energy transfer (FRET) efficiency, these changes can be further analyzed in terms of nanodomain/pore size. This paper is a continuation of previous work on this theme. In particular, it is demonstrated how FRET experiments should be planned and how data should be analyzed in order to achieve the best possible resolution. The limiting resolution of domains and pores are discussed simultaneously, in order to enable direct comparison. It appears that choice of suitable donor/acceptor pairs is the most crucial step in the design of experiments. For instance, it is recommended to use DA pairs, which exhibit an increased affinity to pores (i.e., partition coefficients KD,A > 10) for the determination of pore sizes with radii comparable to the Förster radius R0. On the other hand, donors and acceptors exhibiting a high affinity to different phases are better suited for the determination of domain sizes. The experimental setup where donors and acceptors are excluded from the domains/pores should be avoided.
Keywords: FRET; lipid domains; pores; lipid bilayer; rafts; fluorescent probes FRET; lipid domains; pores; lipid bilayer; rafts; fluorescent probes
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Šachl, R.; Johansson, L.B.-Å.; Hof, M. Förster Resonance Energy Transfer (FRET) between Heterogeneously Distributed Probes: Application to Lipid Nanodomains and Pores. Int. J. Mol. Sci. 2012, 13, 16141-16156.

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