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Speckled Microtubules Improve Tracking in Motor-Protein Gliding Assays
| Content Provider | PubMed Central |
|---|---|
| Author | Chisena, Ernest N. Wall, R. Andrew Macosko, Jed C. Holzwarth, George |
| Abstract | Gliding assays of motor proteins such as kinesin, dynein, and myosin are commonly carried out with fluorescently labeled microtubules or filamentous actin. In this paper, we show that speckled microtubules (MTs), prepared by copolymerizing 98% unlabeled tubulin with 2% rhodamine-labeled tubulin, can be localized to ±7.4 nm (24 measurements) in images acquired every 125 ms. If the speckled MTs move at about 800 nm/s, 10 images are sufficient to determine their velocity to a precision of ±6.8 nm/s (6 microtubules, 24 measurements). This velocity precision is 4-fold better than manual methods for measuring the gliding velocity of uniformly labeled MTs by end-point localization. The improved velocity precision will permit the determination of velocity-force curves when 1, 2, and 3 kinesin motors pull a single load in vitro. |
| Related Links | http://dx.doi.org/10.1088/1478-3975/4/1/002 |
| Ending Page | 15 |
| Page Count | 6 |
| Starting Page | 10 |
| File Format | |
| ISSN | 14783975 |
| e-ISSN | 14783975 |
| Journal | Physical biology |
| Issue Number | 1 |
| Volume Number | 4 |
| Language | English |
| Publisher Date | 2007-02-01 |
| Access Restriction | Open |
| Subject Keyword | Biophysics Cell Biology Molecular Biology Structural Biology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Structural Biology Molecular Biology Biophysics |
