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Detection and Quantification of Protein–Microtubules Interactions Using Green Fluorescent Protein Photoconversion
Author(s) -
Brunet Stéphane,
Zimmermann Timo,
Reynaud Emmanuel G.,
Vernos Isabelle,
Karsenti Eric,
Pepperkok Rainer
Publication year - 2006
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/j.1600-0854.2006.00463.x
Subject(s) - biology , green fluorescent protein , microtubule , fluorescence , fluorescent protein , microbiology and biotechnology , protein subcellular localization prediction , protein detection , fluorescence microscope , fluorescent labelling , computational biology , biochemistry , nanotechnology , gene , physics , quantum mechanics , materials science
We present an in vitro system to analyze quantitatively the interactions of green fluorescent protein (GFP)‐tagged recombinant proteins with microtubules. This method relies on photoconversion of GFP and time‐lapse microscopy. Specific interactions can be detected and binding kinetics can be determined rapidly and accurately. This method provides an alternative to classical in vitro microtubule‐binding assays to analyze microtubule‐associated proteins binding to microtubules. It has the potential to be extended to study interactions of proteins or multi‐protein complexes with different biopolymers like actin microfilaments or organelle membranes.