Isolation of intraflagellar transport trains.
Bottom Line: Moreover, the particles forming isolated IFT trains are structurally similar to the individual particles found in the ∼17S gradient peak.Our results provide the first direct evidence that ∼17S particles do indeed compose the IFT trains.The paper also represents the first isolation of the IFT trains, and opens new possibilities for higher resolution studies on their structure and how particles are attached to each other to form the particle trains.
Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.Show MeSH
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Mentions: We next analyzed the material in the gradient fractions at ∼17S, which is expected to consist mainly of isolated particles. After gold-labeling and negative staining, these fractions were shown to contain particles—some of which were labeled—that exhibit a certain degree of structural variability and may be associated with each other into complexes of variable composition (Fig. 8). In order to analyze such structural heterogeneity, we classified labeled particles using a simple and straightforward morphometric approach, i.e., on the basis of the lengths of their longitudinal and transverse axes. Particles sizes clustered into a few discrete peaks (Fig. 9A). Since within each peak particles exhibited a substantial structural homogeneity, not only in size but also in shape (data not shown), we considered this approach to be tenable and functional for our structural analyses.
Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.