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dc.contributor.authorBreitsprecher, Dennis
dc.contributor.authorKiesewetter, Antje K
dc.contributor.authorLinkner, Joern
dc.contributor.authorVinzenz, Marlene
dc.contributor.authorStradal, Theresia E B
dc.contributor.authorSmall, John Victor
dc.contributor.authorCurth, Ute
dc.contributor.authorDickinson, Richard B
dc.contributor.authorFaix, Jan
dc.date.accessioned2015-03-19T09:34:39Zen
dc.date.available2015-03-19T09:34:39Zen
dc.date.issued2011-02-02en
dc.identifier.citationMolecular mechanism of Ena/VASP-mediated actin-filament elongation. 2011, 30 (3):456-67 EMBO J.en
dc.identifier.issn1460-2075en
dc.identifier.pmid21217643en
dc.identifier.doi10.1038/emboj.2010.348en
dc.identifier.urihttp://hdl.handle.net/10033/346880en
dc.description.abstractEna/VASP proteins are implicated in a variety of fundamental cellular processes including axon guidance and cell migration. In vitro, they enhance elongation of actin filaments, but at rates differing in nearly an order of magnitude according to species, raising questions about the molecular determinants of rate control. Chimeras from fast and slow elongating VASP proteins were generated and their ability to promote actin polymerization and to bind G-actin was assessed. By in vitro TIRF microscopy as well as thermodynamic and kinetic analyses, we show that the velocity of VASP-mediated filament elongation depends on G-actin recruitment by the WASP homology 2 motif. Comparison of the experimentally observed elongation rates with a quantitative mathematical model moreover revealed that Ena/VASP-mediated filament elongation displays a saturation dependence on the actin monomer concentration, implying that Ena/VASP proteins, independent of species, are fully saturated with actin in vivo and generally act as potent filament elongators. Moreover, our data showed that spontaneous addition of monomers does not occur during processive VASP-mediated filament elongation on surfaces, suggesting that most filament formation in cells is actively controlled.
dc.language.isoenen
dc.subject.meshActinsen
dc.subject.meshAmino Acid Sequenceen
dc.subject.meshCell Movementen
dc.subject.meshDNA-Binding Proteinsen
dc.subject.meshKineticsen
dc.subject.meshMicroscopy, Fluorescenceen
dc.subject.meshModels, Biologicalen
dc.subject.meshMolecular Sequence Dataen
dc.subject.meshPeptidesen
dc.subject.meshPolymerizationen
dc.subject.meshRecombinant Fusion Proteinsen
dc.subject.meshTime-Lapse Imagingen
dc.titleMolecular mechanism of Ena/VASP-mediated actin-filament elongation.en
dc.typeArticleen
dc.identifier.journalThe EMBO journalen
refterms.dateFOA2018-06-13T01:19:29Z
html.description.abstractEna/VASP proteins are implicated in a variety of fundamental cellular processes including axon guidance and cell migration. In vitro, they enhance elongation of actin filaments, but at rates differing in nearly an order of magnitude according to species, raising questions about the molecular determinants of rate control. Chimeras from fast and slow elongating VASP proteins were generated and their ability to promote actin polymerization and to bind G-actin was assessed. By in vitro TIRF microscopy as well as thermodynamic and kinetic analyses, we show that the velocity of VASP-mediated filament elongation depends on G-actin recruitment by the WASP homology 2 motif. Comparison of the experimentally observed elongation rates with a quantitative mathematical model moreover revealed that Ena/VASP-mediated filament elongation displays a saturation dependence on the actin monomer concentration, implying that Ena/VASP proteins, independent of species, are fully saturated with actin in vivo and generally act as potent filament elongators. Moreover, our data showed that spontaneous addition of monomers does not occur during processive VASP-mediated filament elongation on surfaces, suggesting that most filament formation in cells is actively controlled.


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