title: Detection and quantification of protein-microtubules interactions using green fluorescent protein photoconversion.
abstract: 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.
authors.affiliation: Biologie du Développement, CNRS UMR 7622 bat C-5ème, 9, Quai St Bernard, 75252 Paris Cedex 05, France. sbrunet@snv.jussieu.fr
authors.forename: Stéphane
authors.initials: S
authors.lastname: Brunet
authors.forename: Timo
authors.initials: T
authors.lastname: Zimmermann
authors.forename: Emmanuel G
authors.initials: EG
authors.lastname: Reynaud
authors.forename: Isabelle
authors.initials: I
authors.lastname: Vernos
authors.forename: Eric
authors.initials: E
authors.lastname: Karsenti
authors.forename: Rainer
authors.initials: R
authors.lastname: Pepperkok
mesh_terms: D049452:Green Fluorescent Proteins; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D008869:Microtubule-Associated Proteins; D049452:Green Fluorescent Proteins