Abstract
Incubation of tubulin with taxol resulted in the assembly of tubulin in the absence of microtubule associated proteins. Optimum assembly occurred at approximately equal concentrations of tubulin and taxol. Both podophyllotoxin and colchicine inhibited the taxol-induced tubulin polymerization. Microtubules formed with taxol were also resistant to podophyllotoxin-induced depolymerization. The rate of tubulin subunit exchange into taxol-induced microtubules at steady state was 5-fold lower than into microtubule-associated protein (MAP2)-stimulated microtubules. There was a marked difference in the kinetics of tubulin polymerized in the presence of both taxol and MAP2 as compared to that obtained with either of them alone. In binding experiments, no competition was observed between taxol and MAP2 or the anti-tubulin drugs, e.g. colchicine, podophyllotoxin, and vinblastine.
Highlights
Incubation of tubulin with taxol resulted in the as- of tubulin in the absenceof MAPS.’ Several experimentsalso sembly of tubulin in the absence of microtubule asso- suggest that taxol does not affect the binding of colchicine, ciatedproteins
Optimum assemblyoccurred at ap- vinblastine, podophyllotoxin, and MAP2 to tubulin. proximately equal concentrations of tubulin atnadxol
Of tubulin was recorded and thefirst order kinetic analysisof the data is shown in Fig. 7B.The kinetic patterns were similar for bothsamples andresembled that obtained withthree-cycle-purified tubulin + taxol (Fig. 7 B ) and not + with PC tubulin + MAP2 taxol (Fig. 7A)
Summary
From the Section on OrganelleBiochemistry, Laboratory of Cell Biology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland20205. Incubation of tubulin with taxol resulted in the as- of tubulin in the absenceof MAPS.’. Several experimentsalso sembly of tubulin in the absence of microtubule asso- suggest that taxol does not affect the binding of colchicine, ciatedproteins. Optimum assemblyoccurred at ap- vinblastine, podophyllotoxin, and MAP2 to tubulin. Optimum assemblyoccurred at ap- vinblastine, podophyllotoxin, and MAP2 to tubulin. proximately equal concentrations of tubulin atnadxol
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