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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Miyamae, M., Camacho, S. A., Weiner, M. W., and Figueredo, V. M. (1996) Alcohol consumption reduces ischemia-reperfusion injury by species-specific signaling in guinea pigs and rats. Am. J. Physiol. 271, H1245–H1253.
Das, D. K. and Maulik, N. (1994) Antioxidant effectiveness in ischemia-reperfusion tissue injury. Methods Enzymol. 233, 601–610.
Chien, K. R., Reeves, J. P., Buja, L. M., Bonte, F., Parkey, R. W., and Willerson, J. T. (1981) Phospholipid alterations in canine ischemic myocardium. Temporal and topographical correlations with Tc-99m-PPi accumulation and an in vitro sarcolemmal Ca2+ permeability defect. Circ. Res. 48, 711–717.
Unlu, M., Morgan, M. E., and Minden, J. S. (1997) Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. Electrophoresis 18, 2071–2077.
Alban, A., David, S. O., Bjorkesten, L., et al. (2003) A novel experimental design for comparative two-dimensional gel analysis: two-dimensional difference gel electrophoresis incorporating a pooled internal standard. Proteomics 3, 36–44.
Shaw, J., Rowlinson, R., Nickson, J., et al. (2003) Evaluation of saturation labeling two-dimensional difference gel electrophoresis fluorescent dyes. Proteomics 3, 1181–1195.
Yamamoto, S., Matsui, K., Kitano, M., and Ohashi, N. (2000) SM-20550, a new Na+/H+ exchange inhibitor and its cardioprotective effect in ischemic/reperfused isolated rat hearts by preventing Ca2+-overload. J. Cardiovasc. Pharmacol. 35, 855–862.
Mizukami, Y., Hirata, T., and Yoshida, K. (1997) Nuclear translocation of PKC zeta during ischemia and its inhibition by wortmannin, an inhibitor of phosphatidylinositol 3-kinase. FEBS Lett. 401, 247–251.
Yoshida, K., Hirata, T., Akita, Y., et al. (1996) Translocation of protein kinase C-alpha, delta and epsilon isoforms in ischemic rat heart. Biochim. Biophys. Acta 1317, 36–44.
Albert, C. J. and Ford, D. A. (1999) Protein kinase C translocation and PKC-dependent protein phosphorylation during myocardial ischemia. Am. J. Physiol. 276(2 Pt 2), H642–650.
Gordon, J. A. (1991) Use of vanadate as protein-phosphotyrosine phosphatase inhibitor. Methods Enzymol. 201, 477–482.
Sakai, J., Ishikawa, H., Kojima, S., Satoh, H., Yamamoto, S., and Kanaoka, M. (2003) Proteomic analysis of rat heart in ischemia and ischemia-reperfusion using fluorescence two-dimensional difference gel electrophoresis. Proteomics 3, 1318–1324.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
Sakai, J., Ishikawa, H., Satoh, H., Yamamoto, S., Kojima, S., Kanaoka, M. (2007). Two-Dimensional Differential Gel Electrophoresis of Rat Heart Proteins in Ischemia and Ischemia-Reperfusion. In: Vivanco, F. (eds) Cardiovascular Proteomics. Methods in Molecular Biology™, vol 357. Humana Press. https://doi.org/10.1385/1-59745-214-9:33
Download citation
DOI: https://doi.org/10.1385/1-59745-214-9:33
Publisher Name: Humana Press
Print ISBN: 978-1-58829-535-4
Online ISBN: 978-1-59745-214-4
eBook Packages: Springer Protocols