Schneider, C., Sepp Lorenzino, L., Nimmesgern, E., Ouerfelli, O., Danishefsky, S., Rosen, N., Hartl, F. U.
Notes: In this report, Hsp90, in cooperation with Hsc70, p60 and other factors, was shown to mediate the refolding of thermally denatured proteins, such as firefly luciferase, both in vivo and in vitro. Luciferase-myc-His, generated in E. coli, was placed in desalted Nuclease-Treated Rabbit Reticulocyte Lysate System (RRL), or luciferase was translated in the RRL. Luciferase:chaperone complexes were generated either by adding the luciferase-myc-His to the RRL and treating with apyrase or by incubation of native luciferase in the RRL followed by treatment with apyrase. The complexes were immunoisolated, and the proteins in them were characterized, including Hip, Hsp40 and p23. Incubation of lysates with geldanamycin and herbimycin A indicated that Hsp90 plays a role in ATP-dependent luciferase processing. (0414)