微小細管の再構成について

概要

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Quite recently, Stephens (1968) reported a success in reassociation of microtubule protein. We have confirmed his results both by measuring flow birefringence and by electron microscopy, using negative staining technique. Microtubules were isolated from sperm flagella of sea-urchin, Anthocidaris crassispina, as described previously (Mohri, 1968), and were depolymerized with 0.5% sodium lauroyl sarcosine. The supernatant after centrifugation for 15 min. at 35,000×g, containing few of fibrous materials, was diluted 10 times with deionized water. The dilution resulted in a formation of "ribbons", 1-3μ in length, consisting of 3-4 arrays of small globular units. The addition of 0.1M KCl caused elongation, up to several μ, and widening of "ribbons", the appearance of which resembles that of native microtubules. Interpretation of electron micrographs, however, seems to favor side-by-side connection of a pair of "ribbons" rather than reconstruction of real microtubules. If the centrifugation was carried out for 1 hr.at 100,000×g, the supernatant freed of any fibrous structures lost the ability to reassociate into "ribbons". The addition of sonicated fragments of microtubule as "seed" to the dilution medium for the 100,000×g supernatant resulted in polymerization of the fragments to some extent.
社団法人日本動物学会の論文
1968-12-15