Recombinant human cytoplasmic dynein heavy chain 1 and 2: Observation of dynein-2 motor activity in vitro

► Expression systems for human cytoplasmic heavy chains of dynein-1 and dynein-2. ► Structural difference between two cytoplasmic dynein isoforms. ► Demonstration of the minus-end-directed microtubule motor activity of dynein-2. Cytoplasmic dynein is a microtubule (MT) motor protein comprising two c...

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Published inFEBS letters Vol. 585; no. 15; pp. 2419 - 2423
Main Authors Ichikawa, Muneyoshi, Watanabe, Yuta, Murayama, Takashi, Toyoshima, Yoko Yano
Format Journal Article
LanguageEnglish
Published England Elsevier B.V 04.08.2011
Subjects
Biological Transport
Cell Line
Cytoplasmic dynein-1
Cytoplasmic dynein-2
Cytoplasmic Dyneins - metabolism
Flagella - metabolism
Humans
IFT
In vitro motility
Intraflagellar transport
Microscopy, Electron
Microtubule
Movement
Recombinant Proteins
SBP
streptavidin-binding peptide
SBP
Cytoplasmic dynein-2
Cytoplasmic dynein-1
Intraflagellar transport
IFT
In vitro motility
Microtubule
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Summary:► Expression systems for human cytoplasmic heavy chains of dynein-1 and dynein-2. ► Structural difference between two cytoplasmic dynein isoforms. ► Demonstration of the minus-end-directed microtubule motor activity of dynein-2. 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. DYNC1H1 (Homo sapiens)binds to DYNC1H1 (Homo sapiens) by electron microscopy(View interaction) DYNC1H1 (Sus scrofa)binds to DYNC1H1 (Sus scrofa) by electron microscopy(View interaction) DYNC2H1 (Homo sapiens)binds to DYNC2H1 (Homo sapiens) by electron microscopy(View interaction)
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ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2011.06.026