- Western Blot (and immunohistochemistry)
Human, mouse and rat acinus
Synthesized S422 phosphorylated peptide (CRSRS(PO3)RDRRRKERAKS-COOH) was conjugated to KLH and injected into rabbits. Antibodies were affinity purified from serum by use of unphosphorylated peptide crosslinked to Affi-Gel 10 or Affi-Gel 15.
150-200 mL at 1.5-3 mg/mL
Acinus, predominantly located in the nucleus, induces apoptotic chromatin condensation after cleavage by caspases. Dr. Ye's group demonstrated that Akt phosphorylates acinus at both S422 and 573 residues, and prevents its proteolytic cleavage in vitro and in vivo. The protective effect is predominantly associated with S422, but not 573 phosphorylation. Cells transfected with acinus (S422, 573A) mutant, unable to be phosphorylated by Akt, reveal robust apoptotic degradation of acinus and chromatin condensation, which are prevented by acinus (S422, 573D), a mutant mimicking phosphorylation. These findings point to acinus as a key substrate for Akt and demonstrate a novel mechanism by which nuclear Akt controls chromatin condensation and apoptosis.
Publication: Hu et. al., (2005). EMBO, J. 24:3543-3554.