Figure 4
Lysine-270 and lysine-277 substitutions convert Hsp90β to Hsp90α-like protein to promote cancer cell motility and invasion. (A) A schematic representation of lysine-270 and lysine-277 in full-length Hsp90α and creations of the Hsp90α-K270G/K277T and Hsp90α -D271K mutants. (B) A schematic representation of glycine-262 and threonine-269 in full-length Hsp90β and creations of the Hsp90β-G262K/T269K and Hsp90β-K263D mutants. (C) Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) with Coomassie Brilliant Blue staining of FPLC-purified recombinant Hsp90α and Hsp90β proteins. Commercial bovine serum albumin (BSA) is included as a comparison for relative quantities. (D) Circular dichroism (CD) revealed the secondary structural profiles of Hsp90α, Hsp90β, and their mutant proteins. (E) Rescue of the invasion defect of the KO-α-#1 cells (panel b vs panel a) by supplementation with wild-type Hsp90α (panel c), Hsp90α-G/T mutant (panel d), Hsp90α-D271K mutant (panel e), wild-type Hsp90β (panel f), Hsp90βK/K mutant (panel g) and Hsp90β-K263D mutant (panel h) (30 μg/ml). Quantitation of invasion (Inv. %) is included.
