Reconstitution of flagellar sliding

Methods Enzymol. 2013;524:343-69. doi: 10.1016/B978-0-12-397945-2.00019-6.

Abstract

The motile structure within eukaryotic cilia and flagella is the axoneme. This structure typically consists of nine doublet microtubules arranged around a pair of singlet microtubules. The axoneme contains more than 650 different proteins that have structural, force-generating, and regulatory functions. Early studies on sea urchin sperm identified the force-generating components, the dynein motors. It was shown that dynein can slide adjacent doublet microtubules in the presence of ATP. How this sliding gives rise to the beating of the axoneme is still unknown. Reconstitution assays provide a clean system, free from cellular effects, to elucidate the underlying beating mechanisms. These assays can be used to identify the components that are both necessary and sufficient for the generation of flagellar beating.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Axonemal Dyneins / isolation & purification
  • Axonemal Dyneins / metabolism*
  • Axoneme / chemistry
  • Axoneme / metabolism*
  • Cell Culture Techniques
  • Cell Engineering
  • Chlamydomonas reinhardtii / chemistry
  • Chlamydomonas reinhardtii / metabolism*
  • Culture Media
  • Flagella / chemistry
  • Flagella / metabolism*
  • Kinesin / metabolism*
  • Kymography
  • Microscopy, Fluorescence
  • Molecular Imaging
  • Movement
  • Tubulin / isolation & purification
  • Tubulin / metabolism*

Substances

  • Culture Media
  • Tubulin
  • Adenosine Triphosphate
  • Axonemal Dyneins
  • Kinesin