ArticleMethods in enzymology2011
Changepoint analysis for single-molecule polarized total internal reflection fluorescence microscopy experiments.
Article in Methods in enzymology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Hidden Markov model analysis of fluorescence blinking in fluorescently labeled DNA.Scientific reports · 2026Article
- The kinesin-5 tail and bipolar minifilament domains are the origin of its microtubule crosslinking and sliding activity.Molecular biology of the cell · 2023Article
- Computational Tool for Ensemble Averaging of Single-Molecule Data.Biophysical journal · 2021Article
- FRET and optical trapping reveal mechanisms of actin activation of the power stroke and phosphate release in myosin V.The Journal of biological chemistry · 2020Article
- ALS/FTLD-Linked Mutations in FUS Glycine Residues Cause Accelerated Gelation and Reduced Interactions with Wild-Type FUS.Molecular cell · 2020Article
- Life under the Microscope: Single-Molecule Fluorescence Highlights the RNA World.Chemical reviews · 2018Review
- Angular measurements of the dynein ring reveal a stepping mechanism dependent on a flexible stalk.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Elongation factor G initiates translocation through a power stroke.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Breaking the millisecond barrier: single molecule motors wobble to find their next binding sites.Biophysical journal · 2013Article
- Tilting and wobble of myosin V by high-speed single-molecule polarized fluorescence microscopy.Biophysical journal · 2013Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
The experimental study of individual macromolecules has opened a door to determining the details of their mechanochemical operation. Motor enzymes such as the myosin family have been particularly attractive targets for such study, in part because some of them are highly processive and their "product" is spatial motion. But single-molecule resolution comes with its own costs and limitations. Often, the observations rest on single fluorescent dye molecules, which emit a limited number of photons before photobleaching and are subject to complex internal dynamics. Thus, it is important to develop methods that extract the maximum useful information from a finite set of detected photons. We have extended an experimental technique, multiple polarization illumination in total internal reflection fluorescence microscopy (polTIRF), to record the arrival time and polarization state of each individual detected photon. We also extended an analysis technique, previously applied to FRET experiments, that optimally determines times of changes in photon emission rates. Combining these improvements allows us to identify the structural dynamics of a molecular motor (myosin V) with unprecedented detail and temporal resolution.
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