In Vitro Reconstitution of Molecular Motor-Driven Mitochondrial Transport

Methods Mol Biol. 2022:2431:533-546. doi: 10.1007/978-1-0716-1990-2_28.

Abstract

Intracellular trafficking of organelles driven by molecular motors underlies essential cellular processes. Mitochondria, the powerhouses of the cell, are one of the major cargoes of molecular motors. Efficient distribution of mitochondria ensures cellular fitness while defects in this process contribute to severe pathologies, such as neurodegenerative diseases. Reconstitution of the mitochondrial microtubule-based transport in vitro in a bottom-up approach provides a powerful tool to investigate the mitochondrial trafficking machinery in a controlled environment in the absence of complex intracellular interactions. In this chapter, we describe the procedures for achieving such reconstitution of mitochondrial transport.

Keywords: Adaptor proteins; Interference reflection microscopy; Kinesin-1; Mitochondria; Molecular motors; Motility assay; TIRF microscopy; TRAK.

MeSH terms

  • Biological Transport
  • Kinesins*
  • Microtubules* / metabolism
  • Mitochondria / metabolism
  • Organelles

Substances

  • Kinesins