Figure 2.
Mutation of K294 Affects Hsp90 Interaction with Client Proteins
(A) COS7 cells were co-transfected with ErbB2 (+) and indicated FLAG-Hsp90 constructs. Cells were lysed and FLAG-Hsp90 immunoprecipitates were separated by SDS-PAGE; ErbB2 binding was detected by immunoblotting.
(B) SkBr3 cells were transfected with indicated FLAG-Hsp90 constructs. Hsp90 was immunoprecipitated from cell lysates and ErbB2 binding was determined as in (A).
(C) NIH3T3 cells stably transfected with v-src were transfected with indicated FLAG-Hsp90 constructs. P60v-Src was immunoprecipitated.and associated FLAG-Hsp90 was detected by immunoblotting.
(D) SkBr3 cells were transfected with indicated FLAG-Hsp90 constructs. Endogenous mutant p53 was immunoprecipitated and associated FLAG-Hsp90 was detected as in (C).
(E) COS7 cells were co-transfected with HA-HIF-1αand indicated FLAG-Hsp90 constructs. HA-HIF-1αwas immunoprecipitated and associated FLAG-Hsp90 was detected as in (C).
(F) COS7 cells were co-transfected with androgen receptor and indicated FLAG-Hsp90 constructs. FLAG-Hsp90 was immunoprecipitated and androgen receptor was detected by immunoblotting.