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Cardiovascular Proteomics

Volume 357 of the series Methods in Molecular Biology™ pp 33-43

Two-Dimensional Differential Gel Electrophoresis of Rat Heart Proteins in Ischemia and Ischemia-Reperfusion

  • Jun SakaiAffiliated withGenomic Science Laboratories, Sumitomo Pharmaceuticals
  • , Hironori IshikawaAffiliated withGenomic Science Laboratories, Sumitomo Pharmaceuticals
  • , Hideshi SatohAffiliated withDiscovery Research Laboratories I, Sumitomo Pharmaceuticals
  • , Setsuko YamamotoAffiliated withDiscovery Research Laboratories I, Sumitomo Pharmaceuticals
  • , Shinichi KojimaAffiliated withGenomic Science Laboratories, Sumitomo Pharmaceuticals
  • , Masaharu KanaokaAffiliated withGenomic Science Laboratories, Sumitomo Pharmaceuticals

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Abstract

Ischemia-reperfusion injury occurs in acute myocardial infarction, cardiopulmonary bypass surgery, and heart transplantation. However the precise mechanisms still remain unclear. In order to identify proteins that are involved in ischemia-reperfusion injury, we compared precipitated 100,000g fractions of normal, ischemic, and ischemic-reperfused rat hearts using two-dimensional (2D) difference gel electrophoresis (2D-DIGE). 2D-DIGE is reliable method to define quantitative protein differences, especially when subtle protein changes are under investigation. In this study, six spots that changed more than twofold and two additional spots related to these spots were detected. Five of the spots were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry as protein disulfide isomerase, one as 60 kDa heat-shock protein, and two as elongation factor Tu.

Key Words

Ischemia ischemia-reperfusion 2D-DIGE PDA3 HSP60 EF-Tu