| Autor: |
Jagoda Rokicka, Ronald S. Rock, Agata K. Krenc |
| Rok vydání: |
2016 |
| Předmět: |
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| Zdroj: |
Biophysical Journal. 110:616a |
| ISSN: |
0006-3495 |
| DOI: |
10.1016/j.bpj.2015.11.3305 |
| Popis: |
Further development of an accurate model of actomyosin network function requires detailed knowledge of the interaction between actin and myosin. The majority of data demonstrating the function of myosins has been obtained either in a single molecule fashion (where the isolated motors have been studied) or in a gliding filament assay (where multiple motors work collectively). Here, I present a technique that joins collective motor mechanics with single molecule detection of binding events, using Fluorescence Interference Contrast Microscopy (FLIC).In the FLIC assay, the distance between a fluorescently labeled actin filament and a reflective surface of a slide can be determined. The locations of myosin molecules that tether the actin filament to the surface are clearly visible. An attachment lifetime of each actin-motor interaction can be easily extracted from these data. The length of actin between two myosins (buckled or under tension) provides information about the relative speed of the two myosins. Moreover, segments of actin under tension and compression transduce different amounts of force to the myosins at each end.Our data show that the attachment lifetimes of Myosin 6 and Non-Muscle Myosin IIB are much longer in gliding filament assay than the attachment lifetimes previously obtained in single molecule bead assay. We argue that this difference, at least partially, can be attributed to mechanical coupling through the actin filament. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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