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Fluorescence Depolarisation Studies of Filamentous Actin Analysed with a Genetic Algorithm
Umeå University, Faculty of Science and Technology, Chemistry.
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2007 (English)In: Biophysical Journal, Vol. 93, 3291-99 p.Article in journal (Refereed) Published
Abstract [en]

A new method, in which a genetic algorithm (GA) was combined with Brownian dynamics and Monte Carlo simulations, was developed to analyse fluorescence depolarisation data collected by the time-correlated single photon counting technique. It was applied to studies of BODIPY-{N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl}-labelled filamentous actin (F-actin). The technique registered the local order and reorienting motions of the fluorophores, which were covalently coupled to cysteine 374 (C374) in actin and interacted by electronic energy migration within the actin polymers. Analyses of F-actin samples composed of different fractions of labelled actin molecules revealed the known helical organization of F-actin, demonstrating the usefulness of this technique for structure determination of complex protein polymers. The distance from the filament axis to the fluorophore was found to be considerably less than expected from the proposed position of C374 at a high filament radius. In addition, polymerisation experiments with BODIPY-actin suggest a 25-fold more efficient signal for filament formation than pyrene-actin.

Place, publisher, year, edition, pages
2007. Vol. 93, 3291-99 p.
Keyword [en]
BODIPY, Genetic algorithm, actin polymerisation, donor-donor energy migration, homotransfer, protein polymer structures
URN: urn:nbn:se:umu:diva-16020DOI: doi:10.1529/biophysj.107.107920OAI: diva2:155693
Available from: 2007-10-16 Created: 2007-10-16 Last updated: 2011-01-11Bibliographically approved

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