Single molecule FRET observation of kinesin-1's head-tail interaction on microtubule

• Aoki Takahiro

  Department of Applied Physics, School of Engineering, the University of Tokyo

• Tomishige Michio

  Department of Applied Physics, School of Engineering, the University of Tokyo

• Ariga Takayuki

  Department of Applied Physics, School of Engineering, the University of Tokyo

Abstract

Kinesin-1 (conventional kinesin) is a molecular motor that transports various cargo such as endoplasmic reticulum and mitochondria in cells. Its two head domains walk along microtubule by hydrolyzing ATP, while the tail domains at the end of the long stalk bind to the cargo. When a kinesin is not carrying cargo, its motility and ATPase activity is inhibited by direct interactions between the tail and head. However, the mechanism of this tail regulation is not well understood. Here, we apply single molecule fluorescence resonance energy transfer (smFRET) to observe this interaction in stalk-truncated kinesin. We found that kinesin with two tails forms a folding conformation and dissociates from microtubules, whereas kinesin with one tail remains bound to the microtubule and is immobile even in the presence of ATP. We further investigated the head-tail interaction as well as head-head coordination on the microtubule at various nucleotide conditions. From these results, we propose a two-step inhibition model for kinesin motility.<br>

Journal

• BIOPHYSICS

  BIOPHYSICS 9 (0), 149-159, 2013

  The Biophysical Society of Japan

Keywords

• molecular motor
• smFRET
• autoinhibition
• tail-regulation
• cargo transport

Details

• CRID: 1390282680199439872
• NII Article ID: 130003383784
• DOI: 10.2142/biophysics.9.149
• ISSN: 13492942
• Web Site: https://www.jstage.jst.go.jp/article/biophysics/9/0/9_149/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed