Fluorescence fluctuations of quantum-dot sensors capture intracellular protein interaction dynamics.
We extend the in vitro principle of co-immunoprecipitation to quantify dynamic protein interactions in living cells. Using a multiresolution implementation of fluorescence correlation spectroscopy to achieve maximal temporal resolution, we monitored the interactions of endogenous bait proteins, recruited by quantum dots, with fluorescently tagged prey. With this approach, we analyzed the rapid physiological regulation of protein kinase A.
Zamir
Eli
E
Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany.
Lommerse
Piet H M
PH
Kinkhabwala
Ali
A
Grecco
Hernán E
HE
Bastiaens
Philippe I H
PI
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D047468:Immunoprecipitation / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods*; D045663:Quantum Dots; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases