title: Recombinant human cytoplasmic dynein heavy chain 1 and 2: observation of dynein-2 motor activity in vitro.
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.
authors: Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
authors: Muneyoshi
authors: M
authors: Ichikawa
authors: Yuta
authors: Y
authors: Watanabe
authors: Takashi
authors: T
authors: Murayama
authors: Yoko Yano
authors: YY
authors: Toyoshima
mesh_terms: D001692:Biological Transport; D002460:Cell Line; D056445:Cytoplasmic Dyneins / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism; D006801:Humans; D008854:Microscopy, Electron; D009068:Movement; D011994:Recombinant Proteins
chemical_list: C511240:DYNC1H1 protein, human; C099883:DYNC2H1 protein, human; D011994:Recombinant Proteins; D056445:Cytoplasmic Dyneins