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Journal of Biomedical Optics

Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging
Author(s): Christoph Biskup; Laimonas Kelbauskas; Thomas Zimmer; Klaus Benndorf; Axel Bergmann; Wolfgang Becker; J. Peter Ruppersberg; Clemens Stockklausner; Nikolaj Klöcker
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Paper Abstract

Resonance energy transfer (RET) has been extensively used to estimate the distance between two different fluorophores. This study demonstrates how protein–protein interactions can be visualized and quantified in living cells by time-correlated single-photon counting (TCSPC) imaging techniques that exploit the RET between appropriate fluorescent labels. We used this method to investigate the association of the potassium inward rectifier channel Kir2.1 and the neuronal PDZ protein PSD-95, which has been implicated in subcellular targeting and clustering of ion channels. Our data show that the two proteins not only colocalize within clusters but also interact with each other. Moreover, the data allow a spatially resolved quantification of this protein–protein interaction with respect to the relative number and the proximity between interacting molecules. Depending on the subcellular localization, a fraction of 20 to 60% of PSD-95 molecules interacted with Kir2.1 channels, approximating their fluorescent labels by less than 5 nm.

Paper Details

Date Published: 1 July 2004
PDF: 7 pages
J. Biomed. Opt. 9(4) doi: 10.1117/1.1755721
Published in: Journal of Biomedical Optics Volume 9, Issue 4
Show Author Affiliations
Christoph Biskup, Friedrich-Schiller-Univ. Jena (Germany)
Laimonas Kelbauskas, Fredrich-Schiller Univ. Jena (Germany)
Thomas Zimmer, Friedrich-Schiller-Univ. Jena (Germany)
Klaus Benndorf, Friedrich-Schiller-Univ. (Germany)
Axel Bergmann, Becker and Hickl GmbH (Germany)
Wolfgang Becker, Becker & Hickl GmbH (Germany)
J. Peter Ruppersberg, SEED GmbH (Germany)
Clemens Stockklausner, Albert-Ludwigs Univ. Freiburg (Germany)
Nikolaj Klöcker, Albert-Ludwigs Univ. Freiburg (Germany)

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