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J. Biol. Chem., Vol. 265, Issue 3, 1639-1647, 01, 1990

Microtubule solutions display nematic liquid crystalline structure

AL Hitt, AR Cross and RC Williams Jr
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235.

We report a study of the spontaneous formation of ordered arrays of microtubules in solution. Form birefringence and anisotropic light- scattering appear rapidly and spontaneously when tubulin, initially present in homogeneous solution, self-assembles into microtubules. This phenomenon is reversible and occurs at protein concentrations of a few milligrams per ml, in the presence or absence of microtubule-associated proteins. Light and electron microscopic examination reveals that extensive regions of these birefringent solutions consist of nearly parallel microtubules. Measurement of the order parameter, S, yields a value of 0.81 +/- 0.05, indicating a high degree of alignment. Comparison of these observations to qualitative predictions developed from the theory of Onsager ((1949) Ann. N.Y. Acad. Sci. 51, 627-659) leads to the conclusion that microtubules form a nematic liquid crystalline phase in vitro under ordinary conditions. Simultaneous spectrophotometric observation of turbidity (a measure of microtubule assembly) and birefringence shows that the parallel ordering lags only slightly behind assembly, thus demonstrating that much microtubule growth must occur by addition of tubulin to the ends of microtubules that are already aligned. These observations of anisotropy are important to the understanding of microtubule dynamics in vitro.
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