Comparative phosphoproteome analysis of cardiomyocytes preconditioned by diazoxide

AIM: To analyze and identify the phosphoproteins associated with diazoxide preconditioning. METHODS: Proteomics technique was used to investigate the changes of phosphoprotein after diazoxide preconditioning. Adult rat ventricular myocytes were pretreated in the presence and absence of 200 μmol/L diazoxide for 10 min. Phosphoproteins prepared and enriched respectively from control and diazoxide pretreated groups were then separated by two-dimensional (2D) gel electrophoresis and stained with sliver staining kit. Phosphoproteins of interest were further identified by mass spectrometry. RESULTS: Associated with diazoxide preconditioning, the proteins of chaperonin containing TCP-1 and hypothetical protein XP_346548 were phosphorylated significantly. The proteins of 94 kD glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphorylated markedly (P<0.05). CONCLUSION: These findings suggest that cardiomyocytes undergo significant posttranslational modification via phosphorylation in a multitude of proteins in response to diazoxide preconditioned signaling, which may mediate myocardioprotection signaling downstream mitochondrial staining dish K_ATP channel induced by ischemic preconditioning.