Single-Molecule Studies on the Motion and Force Generation of the Kinesin-3 Motor KIF1A

Research output: Chapter in Book/Report/Conference proceedingChapter


KIF1A is a neuron-specific member of the kinesin-3 family of microtubule (MT) plus-end-directed motor proteins. It powers the migration of nuclei in differentiating brain stem cells and the transport of synaptic precursors and dense core vesicles in axons. Its dysfunction causes severe neurodevelopmental and neurodegenerative diseases termed KIF1A-associated neurological disorders (KAND). KAND mutations span the entirety of the KIF1A protein sequence, of which the majority are located within the motor domain and are thus predicted to affect the motor’s motility and force-generating properties. Unfortunately, the molecular etiologies of KAND remain poorly understood, in part because KIF1A’s molecular mechanism remains unclear. Here, we describe detailed methods for how to express a tail-truncated dimeric KIF1A in E. coli cells and provide step-by-step protocols for performing single-molecule studies with total internal reflection fluorescence microscopy and optical tweezers assays, which, when combined with structure-function studies, help to decipher KIF1A’s molecular mechanism.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages24
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • E. coli
  • His-tag purification
  • KIF1A
  • Kinesin
  • Molecular cloning
  • Optical tweezers
  • Recombinant protein expression
  • Single-molecule
  • TIRF

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


Dive into the research topics of 'Single-Molecule Studies on the Motion and Force Generation of the Kinesin-3 Motor KIF1A'. Together they form a unique fingerprint.

Cite this