FIG. 5.

Bulk interactions of Hip forms and their association with an Hsp70 mutant lacking ATPase activity. Recombinant wtHip or APAV² was preadsorbed to a covalently coupled immunoaffinity resin. The resins were subsequently incubated in RL containing an ATP-regenerating system. Proteins on the resin complexes were separated by SDS-PAGE and Coomassie blue stained (upper panel). Stained bands representing coprecipitating Hsp70, Hop, and Hsp90 are indicated on the left. Radiolabeled Hsp70 forms were included in the bulk reaction mixtures, and the association of these forms with wtHip and APAV² was resolved by autoradiography of the stained gel (lower panel). Either radiolabeled wild-type Hsp70 (wt) or an ATPase-deficient point mutant (K71E) was included in the reaction mixtures.