Differentially expressed phosphoproteins in diazoxide-pretreated ventricular myocytes by two-dimensional electrophoresis and mass spectrometry in vitro  

作　　者：李洪 肖颖彬 高玉琪 杨天德 

机构地区：[1]Departments of Anesthesiology,Xinqiao Hospital,Third Military Medical University,Chongqing 400037,China [2]Department of Pathophysiology,College of High Altitude Military Medicine,Third Military Medical University,Chongqing 400038,China

出　　处：《Journal of Medical Colleges of PLA(China)》2006年第3期143-147,共5页中国人民解放军军医大学学报（英文版）

基　　金：Supported by the National Natural Science Foundation of China (No. 30200089 and No. 30500211)

摘　　要：Objectives To analyze and identify differentially expressed phosphoproteins associated with mitochondrial KATP channel opening. Methods： Adult rat ventricular myocytes were isolated, cultured, and identified, and pretreated without or with 100 μmol/L diazoxide for 10 min. Phosphoproteins prepared and enriched from the control and diazoxide-pretreated cells were separated by two-dimensional gel electrophoresis （2-DE） followed by sliver staining. The obtained interesting phosphoproteins 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 （P〈0. 01）, while the 94-kDa glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphorylated markedly （P〈0. 01）. Conclusion： These findings suggest that cardiomyocytes undergo significant posttranslational modification via phosphorylation in a multitude of proteins in order to respond diazoxide preconditioning, and these phosphorylated protein may mediate the downstream signaling of cardioprotection by mitochondrial KATp channel opening induced by ischemic preconditioning.Objective: To analyze and identify differentially expressed phosphoproteins associated with mitochondrial KATP channel opening. Methods: Adult rat ventricular myocytes were isolated, cultured, and identified, and pretreated without or with 100μmol/L diazoxide for 10 min. Phosphoproteins prepared and enriched from the control and diazoxide-pretreated cells were separated by two-dimensional gel elec-trophoresis (2-DE) followed by sliver staining. The obtained interesting phosphoproteins were further i-dentified by mass spectrometry. Results: Associated with diazoxide preconditioning, the proteins of chap-eronin containing TCP-1 and hypothetical protein XP- 346548 were phosphorylated significantly (P< 0. 01), while the 94-kDa glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphory-lated markedly (P<0. 01). Conclusion: These findings suggest that cardiomyocytes undergo significant posttranslational modification via phosphorylation in a multitude of proteins in order to respond diazoxide preconditioning, and these phosphorylated protein may mediate the downstream signaling of cardioprotec-tion by mitochondrial KATP channel opening induced by ischemic preconditioning.

关 键 词：PRECONDITIONING mitochondrial KATP channel two-dimensional gel electrophoresis ventricular myocytes DIAZOXIDE 

分 类 号：R654[医药卫生—外科学]