dissection of kinesins processivity解剖驱动蛋白的持续.pdfVIP

Dissection of Kinesin’s Processivity 1 1 1 1,2 ¨ 1 Sarah Adio , Johann Jaud , Bettina Ebbing , Matthias Rief , Gunther Woehlke * 1 Physics Department E22, Technical University Munich, Garching, Germany, 2 Munich Center for Integrated Protein Sciences, Munich, Germany Abstract The protein family of kinesins contains processive motor proteins that move stepwise along microtubules. This mechanism requires the precise coupling of the catalytic steps in the two heads, and their precise mechanical coordination. Here we show that these functionalities can be uncoupled in chimera of processive and non-processive kinesins. A chimera with the motor domain of Kinesin-1 and the dimerization domain of a non-processive Kinesin-3 motor behaves qualitatively as conventional kinesin and moves processively in TIRF and bead motility assays, suggesting that spatial proximity of two Kinein-1 motor domains is sufficient for processive behavior. In the reverse chimera, the non-processive motor domains are unable to step along microtubules, despite the presence of the Kinesin-1 neck coiled coil. Still, ATP-binding to one head of these chimera induces ADP-release from the partner head, a characteristic feature of alternating site catalysis. These results show that processive movement of kinesin dimers requires elements in the motor head that respond to ADP-release and induce stepping, in addition to a proper spacing of the motor heads via the neck coiled coil. Citation: Adio S, Jaud J, Ebbing B, Rief M, Woehlke G (2009) Dissection of Kinesin’s Processivity. PLoS ONE 4(2): e4612. doi:10.1371/journal.pone.0004612 Editor: Andreas Hofmann, Griffith University, Australia Received November 7, 2008; Accepted January 21, 2009; Publish