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Measuring microtubule binding kinetics of membrane-bound kinesin motors using supported lipid bilayers.

ABSTRACT: Membrane-bound cargos in cells are generally transported by multiple kinesin motors. Quantifying the bimolecular on-rate of motors for their microtubule track is important for understanding of multi-motor transport but is complicated by diffusion of the motors in the plane of the lipid bilayer. Here, we describe a method to measure the kinesin on-rate that uses a modified microtubule gliding assay performed on a supported lipid bilayer and detects motor binding by a local increase in fluorescence. For complete details on the use and execution of this protocol, please refer to Jiang et al. (2019).

SUBMITTER: Jiang R

PROVIDER: S-EPMC8339383 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Measuring microtubule binding kinetics of membrane-bound kinesin motors using supported lipid bilayers.

Jiang Rui R Hancock William O WO

STAR protocols 20210729 3

Membrane-bound cargos in cells are generally transported by multiple kinesin motors. Quantifying the bimolecular on-rate of motors for their microtubule track is important for understanding of multi-motor transport but is complicated by diffusion of the motors in the plane of the lipid bilayer. Here, we describe a method to measure the kinesin on-rate that uses a modified microtubule gliding assay performed on a supported lipid bilayer and detects motor binding by a local increase in fluorescenc ...[more]

PMID: 34382017

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