AtMAP65-1 Does Not Affect Microtubule Dynamics.
(A) Amount of tubulin sedimented at 100,000g after dilution of the 20-μM tubulin mixture. Squares, tubulin only solution; circles, tubulin with 10 μM AtMAP65-1; triangles, tubulin with 10 μM taxol.
(B) Turbidity of the 20-μM tubulin solution without, and with, the addition of 10 μM AtMAP65-1 (solid lines) at 32°C and after decreasing the temperature. Temperature is indicated by the broken line. The arrow indicates the time at which AtMAP65-1 was added.
(C) Coomassie-stained SDS-PAGE gel of microtubule pellets and supernatants of a 20-μM tubulin solution incubated at 32°C for 10 min, a 20-μM tubulin solution with 10 μM AtMAP65-1 incubated at 32°C for 10 min and then at 1°C for 10 min, and a 20-μM tubulin solution incubated at 32°C for 10 min and then at 1°C for 10 min. The final temperature of the reaction mixtures are indicated below the lanes.