title: The optical trapping dumbbell assay for nonprocessive motors or motors that turn around filaments.
abstract: In vitro motility assays enabled the analysis of coupling between ATP hydrolysis and movement of myosin along actin filaments or kinesin along microtubules. Single-molecule assays using laser trapping have been used to obtain more detailed information about kinesins, myosins, and processive DNA enzymes. The combination of in vitro motility assays with laser-trap measurements has revealed detailed dynamic structural changes associated with the ATPase cycle. This protocol describes the preparation of biotin-actin filaments and coverslips coated with polystyrene beads. These are then used in optical trapping dumbbell assays to study interactions between motors and filaments.
authors: James A
authors: JA
authors: Spudich
authors: Sarah E
authors: SE
authors: Rice
authors: Ronald S
authors: RS
authors: Rock
authors: Thomas J
authors: TJ
authors: Purcell
authors: Hans M
authors: HM
authors: Warrick
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism*; D001710:Biotin / Q000737:chemistry / Q000378:metabolism*; D008863:Microspheres; D020409:Molecular Motor Proteins / Q000378:metabolism*; D052898:Optical Tweezers; D013194:Staining and Labeling
chemical_list: D020409:Molecular Motor Proteins; D001710:Biotin