Proteomic analysis of Rac1 transgenic mice displaying dilated cardiomyopathy reveals an increase in creatine kinase M-chain protein abundance |
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Authors: | Buscemi Nina Doherty-Kirby Amanda Sussman Mark A. Lajoie Gilles Van Eyk Jennifer E. |
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Affiliation: | (1) Department of Physiology, Queen's University, Kingston, Ontario, Canada;(2) Department of Biochemistry, University of Western Ontario, Siebens-Drake Research Institute, London, Ontario, Canada;(3) Division of Molecular and Cardiovascular Biology, The Children's Hospital and Research Foundation, Cincinnati, OH 45229, USA;(4) Departments of Physiology and Biochemistry, Queen's University, Kingston, Ontario, Canada |
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Abstract: | Here, we demonstrate the application of the proteomic approach to the study of a transgenic mouse model of heart failure and provide an example of a disease-associated protein alteration that can be observed using this approach. Specifically, we applied the proteomic approach to the analysis of a mouse model of dilated cardiomyopathy in which the small GTPase, Rac1, was constitutively expressed specifically in the myocardium. We utilized the methods of two-dimensional gel electrophoresis (2-DE) for protein separation, silver-staining for protein visualization and mass spectrometry (MALDI-TOF and MS/MS) for protein spot identification. Computer-generated composite images were created which represent a normalized average of four 2-DE gel images derived from analysis of either Rac1 transgenic (n = 4) or non-transgenic (n = 4) mice. Analysis of composite images derived from NTG and Rac1 experimental groups revealed numerous statistically significant differences in mean protein spot intensities. Here, we report a statistically significant increase, of approximately 1.6-fold, in the mean protein spot intensity for creatine kinase M-chain in the composite image of Rac1 transgenic mice compared to control. This protein alteration may be consistent with an end-stage heart failure phenotype in which maximal myocardial reserve is employed to sustain survival. |
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Keywords: | proteomics heart failure Rac1 |
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