Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging

Journal of Biomedical Optics
C BiskupNikolaj Klöcker

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.

References

Aug 1, 1967·Proceedings of the National Academy of Sciences of the United States of America·L Stryer, R P Haugland
Jun 6, 1998·Nucleic Acids Research·X XuY Luo
May 30, 1998·Cell·S E Craven, D S Bredt
Jun 11, 1999·The Journal of Biological Chemistry·A W McGee, D S Bredt
Jun 17, 2000·Trends in Cell Biology·C C GarnerR L Huganir
Aug 31, 2000·Analytical Biochemistry·G H PattersonB G Barisas
Apr 3, 2001·Annual Review of Neuroscience·M Sheng, C Sala
May 31, 2001·The Journal of Cell Biology·S TomitaD S Bredt
Jan 24, 2002·The Journal of Biological Chemistry·Jeonghoon ChoiEunjoon Kim
Mar 5, 2003·Journal of Molecular Biology·Jia-Fu LongMingjie Zhang
Mar 5, 1965·Journal of the American Chemical Society·S A LATTE R BLOUT

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Citations

Feb 24, 2009·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·L KelbauskasD Lohr
Feb 16, 2008·Immunological Reviews·Raluca A NiesnerMatthias Gunzer
Sep 1, 2011·Microscopy Research and Technique·W BeckerK Weisshart
Feb 22, 2005·Current Opinion in Biotechnology·Horst Wallrabe, Ammasi Periasamy
Jan 25, 2011·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Michael S RobertsWolfgang Becker
Mar 30, 2007·Microscopy Research and Technique·Christoph BiskupKlaus Benndorf
Mar 30, 2007·Microscopy Research and Technique·Wolfgang BeckerChristoph Biskup
Mar 17, 2007·Microscopy Research and Technique·A RückR Steiner
Dec 12, 2007·Chembiochem : a European Journal of Chemical Biology·Sandra OrthausStephan Diekmann
Oct 14, 2010·Optics Express·Raffaella MercatelliFranco Quercioli
May 25, 2012·Journal of Microscopy·W Becker
Jul 9, 2020·Journal of Neuroinflammation·Yuan DongJinbo Cheng
Oct 1, 2004·Journal of Cell Science·Christoph BiskupFrank-D Böhmer

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