Key result
A modified actin sliding filament assay was developed to reduce the number of interacting heads, aiming to characterize the motor function of a single myosin head.
This methodological paper describes an approach to modify in vitro motility assays to study the motor function of single myosin heads.
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In vitro motility assays effectively probe myosin mechanics; leaves open translation to cardiac research and practice.
Over the last five years, the value of in vitro motility assays as probes of the mechanical properties of the actin-myosin interaction has been amply demonstrated. Motility assays in which single fluorescent actin filaments are observed moving over surfaces coated with myosin or its soluble fragments are now used in many laboratories. They have been applied to a wide range of problems including the study of structure-function relationships in the myosin molecule and measurement of fundamental properties of the myosin head. However, one limitation of these assays has been uncertainty over the number of myosin heads interacting with each sliding filament, that frustrates attempts to determine properties of individual heads. In order to address this limitation, we have modified the conditions of the actin sliding filament assay to reduce the number of heads interacting with each filament. Our goal is to establish an assay in which the motor function of a single myosin head can be characterized from the movement of a single actin filament.
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Kron et al. (1991) studied this question. Modified actin sliding filament assay was evaluated on Motor function of a single myosin head. A modified actin sliding filament assay was developed to reduce the number of interacting heads, aiming to characterize the motor function of a single myosin head.
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