Figure 2
Tylophorine analogs interact with Hsc70. (a) Hsc70, but not Hsp70 or Hsp90, bound specifically to rac-cryptopleurine or DCB-3503 affinity resin. (b) Bound recombinant Hsc70 was eluted with increasing concentrations of ATP. The Hsc70 in each fraction was resolved by Western blot. (c) Bound recombinant Hsc70 was eluted with 50 µM ATP or UTP. Hsc70 in each fraction was resolved by Western blot. (d) His-tagged different fragments of Hsc70 were incubated with streptavidin-agarose beads coated with biotinylated-rac-cryptopleurine or -DCB-3503. Bound proteins were immunoblotted with anti-His tag antibody. (e) His-tagged Hsc70 protein harboring D206S mutation was incubated with streptavidin-agarose beads pre-bound to biotinylated-rac-cryptopleurine or -DCB-3503. Bound proteins were immunoblotted with anti-His tag antibody. Results are representative of at least three independent experiments.
