Recombinant Human Cytoplasmic Dynein Heavy Chain 1 and 2: Observation of dynein-2 Motor Activity in Vitro

FEBS Lett. 2011 Aug 4;585(15):2419-23. doi: 10.1016/j.febslet.2011.06.026. Epub 2011 Jun 26.

Abstract

Cytoplasmic dynein is a microtubule (MT) motor protein comprising two classes: dynein-1 and dynein-2. We purified recombinant human dynein-1 and dynein-2 from HEK-293 cells by expressing the streptavidin-binding peptide-tagged human cytoplasmic dynein-1 and dynein-2 heavy chains (HCs), respectively. Electron microscopy of the purified molecules revealed a two-headed structure composed of characteristic dynein motor domains. In an in vitro MT gliding assay, both dynein-1 and dynein-2 showed minus-end-directed motor activities. This is the first demonstration of dynein-2 motor activity, which supports the retrograde intraflagellar transport role of dynein-2. Our expression system of dynein HCs provides a useful means to investigate dynein functions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport
  • Cell Line
  • Cytoplasmic Dyneins / metabolism*
  • Flagella / metabolism
  • Humans
  • Microscopy, Electron
  • Movement
  • Recombinant Proteins

Substances

  • DYNC1H1 protein, human
  • DYNC2H1 protein, human
  • Recombinant Proteins
  • Cytoplasmic Dyneins