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
- 6Citations
- 10Captures
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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
http://www.sciencedirect.com/science/article/pii/S0006291X18318734; http://dx.doi.org/10.1016/j.bbrc.2018.08.178; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=85053005963&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/30213631; https://linkinghub.elsevier.com/retrieve/pii/S0006291X18318734; https://dx.doi.org/10.1016/j.bbrc.2018.08.178
Elsevier BV
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