1 July 2004 Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging
Author Affiliations +
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.
© (2004) Society of Photo-Optical Instrumentation Engineers (SPIE)
Christoph Biskup, Christoph Biskup, Laimonas Kelbauskas, Laimonas Kelbauskas, Thomas Zimmer, Thomas Zimmer, Klaus Benndorf, Klaus Benndorf, Axel Bergmann, Axel Bergmann, Wolfgang Becker, Wolfgang Becker, J. Peter Ruppersberg, J. Peter Ruppersberg, Clemens Stockklausner, Clemens Stockklausner, Nikolaj Klöcker, Nikolaj Klöcker, } "Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging," Journal of Biomedical Optics 9(4), (1 July 2004). https://doi.org/10.1117/1.1755721 . Submission:
JOURNAL ARTICLE
7 PAGES


SHARE
Back to Top