期刊
CYTOSKELETON
卷 73, 期 3, 页码 131-144出版社
WILEY
DOI: 10.1002/cm.21283
关键词
X-ray fiber diffraction analysis; microtubule structure in solution; shear-flow alignment of microtubules; negative thermal expansion of microtubules; GMPCPP; paclitaxel
类别
资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) [20657014, 1704911, 19037010]
- program of Creation and Application of a Soft Nano-Machine
- Hyperfunctional Molecular Machine (CREST, JST)
- Institute of Sciences and Engineering, Chuo University
- Grants-in-Aid for Scientific Research [20657014, 19037010] Funding Source: KAKEN
Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations. (c) 2016 Wiley Periodicals, Inc.
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