Purified tubulin or microtubules are widely used for in vitro assays related to their function, such as activities of modifying enzymes and motor protein activity. This paper gave a review purification (MTs) using two different methods. One method MTs depended on polymerization depolymerization. were polymerized at 37 °C with taxol the presence. Another is purify by affinity column chromatograp...

The dynamic instability of microtubules has long been understood to depend on the hydrolysis of GTP bound to -tubulin, an event stimulated by polymerization and necessary for depolymerization. Crystallographic studies of tubulin show that GTP is bound by -tubulin at the longitudinal dimer-dimer interface and contacts particular -tubulin residues in the next dimer along the protofilament. This s...

Microtubule dynamics and polarity stem from the polymerization of αβ-tubulin heterodimers. Five conserved tubulin cofactors/chaperones and the Arl2 GTPase regulate α- and β-tubulin assembly into heterodimers and maintain the soluble tubulin pool in the cytoplasm, but their physical mechanisms are unknown. Here, we reconstitute a core tubulin chaperone consisting of tubulin cofactors TBCD, TBCE,...

GTP-dependent microtubule polymerization dynamics are required for cell division and are accompanied by domain rearrangements in the polymerizing subunit, -tubulin. Two opposing models describe the role of GTP and its relationship to conformational change in -tubulin. The allosteric model posits that unpolymerized -tubulin adopts a more polymerization-competent conformation upon GTP binding. Th...

A series of derivatives of 5,6-diphenylpyridazin-3-one (DPP) was examined for interactions with calf brain tubulin following the demonstration that many members of the class caused significant mitotic effects in intact animals, while others had activity against murine P388 leukemia. In L1210 cells several DPP derivatives caused a rise in the mitotic index which correlated well with the cytotoxi...

Microtubules are polymers that cycle stochastically between polymerization and depolymerization, i.e. they exhibit "dynamic instability." This behavior is crucial for cell division, motility, and differentiation. Although studies in the last decade have made fundamental breakthroughs in our understanding of how cellular effectors modulate microtubule dynamics, analysis of the relationship betwe...

Several types of non-equilibrium phenomena have been observed in microtubule polymerization, including dynamic instability, assembly overshoot and oscillations. They can be interpreted in terms of interactions between tubulin subunits (= alpha, beta heterodimers), microtubules, and a third state, oligomers, which represent intermediates between microtubule disassembly and the regeneration of as...

The important anticancer drug paclitaxel binds to the beta-subunit of the alphabeta-tubulin dimer in the microtubule in a stoichiometric ratio, promoting microtubule polymerization and stability. The conformation of microtubule-bound drug has been the subject of intense study, and various suggestions have been proposed. In previous work we presented experimental and theoretical evidence that pa...

Assembly-competent tubulin was purified from the cytoplasm of unfertilized and parthogenetically activated oocytes, and from isolated meiotic spindles of the surf clam, Spisula solidissima. At 22 degrees C or 37 degrees C, Spisula tubulin assembled into 48-51-nm macrotubules during the first cycle of polymerization and 25-nm microtubules during the third and subsequent cycles of assembly. Macro...

spinal cord injury (sci) in humans remains a devastating and incurable disorder. a very important obstacle in axonal regeneration after spinal cord injury is astroglial scaring. the use of self-assembling peptide nanofiber, a hydrogel mimicking extracellular matrix, has been suggested as a scaffold for spinal cord regeneration and inhibition of astrogliosis. however, neurogenesis potential of l...