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• W2089367076 abstract "Polarized total internal reflection fluorescence microscopy (polTIRFM) can be used to detect the spatial orientation and rotational dynamics of single molecules. polTIRFM determines the three-dimensional angular orientation and the extent of wobble of a fluorescent probe bound to the macromolecule of interest. This protocol describes the processive motility assay for investigating the motility of myosin V in vitro. Biotin-Alexa actin filaments are fixed to a slide by biotin/streptavidin linkages and aligned with the microscope x-axis by fluid flow. The orientation of a rhodamine-calmodulin (CaM) probe bound to a single myosin V molecule is determined as it moves along an actin filament. Excess wild-type calmodulin (WT-CaM) is present in the buffer solution to replenish lost CaM from the myosin lever arm. The techniques for myosin V should be generally applicable to other single-molecule experiments where angular changes have an important mechanistic role in their biological function." @default.
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• W2089367076 date "2012-06-01" @default.
• W2089367076 modified "2023-09-28" @default.
• W2089367076 title "The Polarized Total Internal Reflection Fluorescence Microscopy (polTIRFM) Processive Motility Assay for Myosin V" @default.
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• W2089367076 doi "https://doi.org/10.1101/pdb.prot069393" @default.
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