Circular orientation fluorescence emitter imaging (COFEI) of rotational motion of motor proteins.
- Resource Type
- Article
- Authors
- Sugawa, Mitsuhiro; Masaike, Tomoko; Mikami, Nagisa; Yamaguchi, Shin; Shibata, Keitaro; Saito, Kei; Fujii, Fumihiko; Toyoshima, Yoko Y.; Nishizaka, Takayuki; Yajima, Junichiro
- Source
- Biochemical & Biophysical Research Communications. Oct2018, Vol. 504 Issue 4, p709-714. 6p.
- Subject
- *FLUORESCENCE
*MOLECULAR motor proteins
*BIOMOLECULES
*NANORODS
*FLUORESCENT probes
- Language
- ISSN
- 0006-291X
Abstract 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. [ABSTRACT FROM AUTHOR]