Defocused orientation and position imaging (DOPI) of myosin V

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Defocused orientation and position imaging (DOPI) of myosin V.

The centroid of a fluorophore can be determined within approximately 1.5-nm accuracy from its focused image through fluorescence imaging with one-nanometer accuracy (FIONA). If, instead, the sample is moved away from the focus, the point-spread-function depends on both the position and 3D orientation of the fluorophore, which can be calculated by defocused orientation and position imaging (DOPI...

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Adaptability of myosin V studied by simultaneous detection of position and orientation.

We studied the structural dynamics of chicken myosin V by combining the localization power of fluorescent imaging with one nanometer accuracy (FIONA) with the ability to detect angular changes of a fluorescent probe. The myosin V was labeled with bifunctional rhodamine on one of its calmodulin light chains. For every 74 nm translocation, the probe exhibited two reorientational motions, associat...

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Accurate Position Sensing of Defocused Beams Using Simulated Beam Templates

In position detection using matched filtering one is faced with the challenge of determining the best position in the presence of distortions such as defocus and diffraction noise. This work evaluates the performance of simulated defocused images as the template against the real defocused beam. It was found that an amplitude modulated phase-only filter is better equipped to deal with real defoc...

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Myosin V

Calcium activates the ATPase activity of tissue-purified myosin V, but not that of shorter expressed constructs. Here, we resolve this discrepancy by comparing an expressed full-length myosin V (dFull) to three shorter constructs. Only dFull has low ATPase activity in EGTA, and significantly higher activity in calcium. Based on hydrodynamic data and electron microscopic images, the inhibited st...

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 2006

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.0507134103