Orientation of actin filaments during motion in in vitro motility assay

J. Borejdo, S. Burlacu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Rhodamine-phalloidin was added to F-actin, and the orientation of transition dipoles of the dye was measured in single actin filaments by polarization of fluorescence. Rhodamine-phalloidin was well immobilized on the surface of actin, indicating that changes in orientation of the dye reported changes in orientation of actin monomers. In stationary filaments the dipoles were inclined at 49.3 degrees with respect to the filament axis. The disorganization of dipoles in stationary filaments was insignificant. When the filaments were made to translate, the average orientation of the dye did not change, but disorganization slightly increased. Disorganization increased significantly when filaments were free in solution. We concluded that, within the accuracy of our measurements (approximately 18%), actin monomers did not undergo major reorientations during motion, but that binding of myosin heads deformed the structure of filaments.

Original languageEnglish
Pages (from-to)1319-1327
Number of pages9
JournalBiophysical Journal
Volume66
Issue number5
DOIs
StatePublished - 1 Jan 1994

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Actin Cytoskeleton
Actins
Coloring Agents
Fluorescence Polarization
Myosins
In Vitro Techniques
rhodamine-phalloidin

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abstract = "Rhodamine-phalloidin was added to F-actin, and the orientation of transition dipoles of the dye was measured in single actin filaments by polarization of fluorescence. Rhodamine-phalloidin was well immobilized on the surface of actin, indicating that changes in orientation of the dye reported changes in orientation of actin monomers. In stationary filaments the dipoles were inclined at 49.3 degrees with respect to the filament axis. The disorganization of dipoles in stationary filaments was insignificant. When the filaments were made to translate, the average orientation of the dye did not change, but disorganization slightly increased. Disorganization increased significantly when filaments were free in solution. We concluded that, within the accuracy of our measurements (approximately 18{\%}), actin monomers did not undergo major reorientations during motion, but that binding of myosin heads deformed the structure of filaments.",
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Orientation of actin filaments during motion in in vitro motility assay. / Borejdo, J.; Burlacu, S.

In: Biophysical Journal, Vol. 66, No. 5, 01.01.1994, p. 1319-1327.

Research output: Contribution to journalArticle

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