Figure 2. Phosphoryaltion of Mesangial Lysate by Recombinant Active Akt.

Candidate Akt substrates were identified by phosphorylation of mesangial lysate with recombinant active Akt in the presence of [γ-P32]ATP. Proteins were separated by 2-DE, and phosphoproteins were detected by autoradiography. Phosphoproteins were identified by comparing autoradiographs with corresponding Sypro-Ruby-stained gels. Five candidate Akt substrates identified by peptide mass fingerprinting of trypsin-digested phosphoproteins using MALDI-MS are indicated by arrows.