The influence of ATP and p23 on the conformation of hsp90.

The chaperoning activity of the heat shock protein hsp90 is directed, in part, by the binding and hydrolysis of ATP and also by association with co-chaperone proteins. One co-chaperone, p23, binds to hsp90 only when hsp90 is in a conformation induced by the binding of ATP. Once formed, the p23-hsp90 complex is very stable upon the removal of ATP and dissipates at 30 degrees with a half-life of about 45 min. This was shown to be due to the high stability of the ATP-induced state of hsp90, not to the rate of p23 dissociation. Further stabilization of this ATP-induced state is achieved by including molybdate or by use of the ATP analogue ATPgammaS. This conformational state of hsp90 is correlated with the tight binding of ADP resulting from hydrolysis of bound ATP. Both p23 and molybdate enhance and stabilize the nucleotide-bound state of hsp90, and this state is maximized by the presence of both agents. These results can be explained in a model where the binding of ATP induces a conformational transition in hsp90 that traps the nucleotide and is committed to ATP hydrolysis. p23 specifically recognizes this state and may also facilitate subsequent steps in the chaperoning cycle.