| Abstract: | The new fluorophor for tubulin, DAPI, is shown to bind to a site different from the exchangeable nucleotide binding site (E site) and to inhibit GTP hydrolysis by the tubulin-colchicine complex within an uncompetitive scheme. Moreover the dissociation rate constant of tubulin for microtubule ends at 32 degrees C was found largely decreased in the presence of saturating amounts of the probe while the association rate constant was little affected. These data on the kinetic parameters of tubulin interactions in the presence of DAPI, together with the inhibition of GTP hydrolysis by microtubules at the steady state are understood as the main cause for microtubule stabilization at steady-state by DAPI. |
