Interaction of Phomopsin A with Normal and Subtilisin-Treated Bovine Brain Tubulin.

Tubulin, the major component of microtubules, has a tendency to lose its ability to assemble or to bind to ligands in a time-dependent process known as “decay.” The decay process also causes tubulin to expose sulfhydryl groups and hydrophobic areas. The antimitotic drug phomopsin A strongly protects the tubulin molecule from decay. Here we have studied the interaction of phomopsin A with αβ tubulin and tubulin which has been treated with subtilisin to remove selectively the C-termini of the α and β chains (α_sβ_s). The binding of phomopsin A to αβ tubulin decreases the sulfhydryl titer by approximately 1.0 mol/mol. Selective removal of the peptides from the C-terminal ends does not affect phomopsin A's interaction with tubulin. Moreover, the α_sβ_s tubulin–phomopsin A complex appears to be more stable than the αβ tubulin–phomopsin A complex as determined by the time-dependent increase in exposure of sulfhydryl groups and hydrophobic areas on tubulin. In fact, phomopsin A inhibits the decay process of α_sβ_s tubulin completely. This observation raises the possibility of determining the conformtion of this configuration of tubulin. [ABSTRACT FROM AUTHOR]