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Circular orientation fluorescence emitter imaging (COFEI) of rotational motion of motor proteins

Biochemical and Biophysical Research Communications, ISSN: 0006-291X, Vol: 504, Issue: 4, Page: 709-714
2018
  • 6
    Citations
  • 0
    Usage
  • 10
    Captures
  • 0
    Mentions
  • 0
    Social Media
Metric Options:   Counts1 Year3 Year

Metrics Details

  • Citations
    6
    • Citation Indexes
      5
    • Policy Citations
      1
      • Policy Citation
        1
  • Captures
    10

Article Description

Single-molecule fluorescence polarization technique has been utilized to detect structural changes in biomolecules and intermolecular interactions. Here we developed a single-molecule fluorescence polarization measurement system, named circular orientation fluorescence emitter imaging (COFEI), in which a ring pattern of an acquired fluorescent image (COFEI image) represents an orientation of a polarization and a polarization factor. Rotation and pattern change of the COFEI image allow us to find changes in the polarization by eye and further values of the parameters of a polarization are determined by simple image analysis with high accuracy. We validated its potential applications of COFEI by three assays: 1) Detection of stepwise rotation of F 1 -ATPase via single quantum nanorod attached to the rotary shaft γ; 2) Visualization of binding of fluorescent ATP analog to the catalytic subunit in F 1 -ATPase; and 3) Association and dissociation of one head of dimeric kinesin-1 on the microtubule during its processive movement through single bifunctional fluorescent probes attached to the head. These results indicate that the COFEI provides us the advantages of the user-friendly measurement system and persuasive data presentations.

Bibliographic Details

Sugawa, Mitsuhiro; Masaike, Tomoko; Mikami, Nagisa; Yamaguchi, Shin; Shibata, Keitaro; Saito, Kei; Fujii, Fumihiko; Toyoshima, Yoko Y; Nishizaka, Takayuki; Yajima, Junichiro

Elsevier BV

Biochemistry, Genetics and Molecular Biology

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