Purification of intact microtubules from brain.
TLDR
Hundred-fold purification of intact microtubules from homogenates of rat brain is reported, and the purified fractions show a major band which migrates like purified tubulin in the SDS gel electrophoresis system.Abstract:Â
Hundred-fold purification of intact microtubules from homogenates of rat brain is reported. The success of purification depends on stabilizing the microtubule structure by the combined effects of hexylene glycol, acidic Ph, and low temperature. A practical, negative stain, electron microscopic assay is used to study purity and stability of microtubule fractions. The purified fractions show a major band which migrates like purified tubulin in the SDS gel electrophoresis system.read more
Citations
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The stabilization of proteins by sucrose.
James Lee,Serge N. Timasheff +1 more
TL;DR: The results from theprotein-solvent interaction study indicate that sucrose is preferentially excluded from the protein domain, increasing the free energy of the system and leading to protein stabilization.
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The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons.
Paul N. Hoffman,Raymond J. Lasek +1 more
TL;DR: The demonstration on SDS gels that the slow component of axonal transport is composed of a small number of polypeptides which have identical molecular weights in neurons from different mammalian species suggests that these polypePTides comprise fundamental structures of vertebrate neurons.
Journal ArticleDOI
Taxol binds to polymerized tubulin in vitro
J. Parness,Susan Band Horwitz +1 more
TL;DR: It is suggested that taxol stabilizes interactions between dimers that lead to microtubule polymer formation and binds specifically and reversibly to a polymerized form of tubulin with a stoichiometry approaching unity.
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The periodic association of MAP2 with brain microtubules in vitro.
TL;DR: A heat-stable fraction highly enriched for one of the MAPs, MAP2, devoid of MAP1, has been purified from calf neurotubules, lowering the critical concentration for tubulin assembly to 0.05 mg/ml and demonstrating a striking axial periodicity.
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Microtubule-associated proteins and the stimulation of tubulin assembly in vitro
TL;DR: MAPs stimulated tubulin assembly by affecting both the initiation and elongation processes, and the effect on initiation was indicated by results showing an increase in initial rate and a decrease in average microtubule length as the MAP:tubulin ratio was increased.
References
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Journal Article
Protein Measurement with the Folin Phenol Reagent
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
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The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis
Klaus Weber,Mary Osborn +1 more
TL;DR: The results show that the polyacrylamide gel electrophoresis method can be used with great confidence to determine the molecular weights of polypeptide chains for a wide variety of proteins.
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Colchicine-binding protein of mammalian brain and its relation to microtubules
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THE MECHANISM OF ACTION OF COLCHICINE Colchicine Binding to Sea Urchin Eggs and the Mitotic Apparatus
Gary G. Borisy,Edwin W. Taylor +1 more
TL;DR: The properties of the colchicine-bindinG protein, (binding constant, sedimentation constant, Sephadex elution volume) are similar to those obtained with the protein from mammalian cells, sea-urchin sperm tails, and brain tissue, and thus support the conclusion that the protein is a subunit of microtubules.
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Regulation of Axon Formation by Clonal Lines of a Neural Tumor
TL;DR: Clonal lines of neuroblastoma cells were found to extend or retract axons depending upon the concentration of serum, suggesting that neurite formation is dependent upon the assembly of microtubules or neurofilaments from preformed protein subunits.