In vitro assembly of pure tubulin into microtubules in the absence of microtubule-associated proteins and glycerol.
Microtubule protein from porcine cerebrum was fractionate into pure tubulin and microtubule-associated proteins by chromatography on phosphocellulose. In agreement with previous studies, pure tubulin does not form microtubules to a significant extent at 37 degrees in normal assembly buffers, which are characterized by a low concentration of Mg2+ ions. If, however, the Mg2+ concentration is raised to approximately 10 mM, rapid and extensive self-assembly of pure tubulin into microtubules is observed, provided the tubulin concentration is above 2.5 mg/ml. At a protein concentration of 3 mg/ml, the lag period is 1.5 min and the assembly process is virtually complete after 6 min at 37 degrees. These microtubules are like normal microtubules--sensitive to calcium ions, colchicine, and low temperature.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002474:Cell-Free System; D003078:Colchicine / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D013552:Swine; D013696:Temperature; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism*
D006023:Glycoproteins; D009419:Nerve Tissue Proteins; D014404:Tubulin; D008274:Magnesium; D005990:Glycerol; D003078:Colchicine; D002118:Calcium
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