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

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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).

Original languageEnglish (US)
Article number100691
JournalSTAR Protocols
Volume2
Issue number3
DOIs
StatePublished - Sep 17 2021

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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