Proteomic analysis of Rac1 transgenic mice displaying dilated cardiomyopathy reveals an increase in creatine kinase M-chain protein abundance.
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.
Department of Physiology, Queen's University, Kingston, Ontario, Canada.
Nina
N
Buscemi
Amanda
A
Doherty-Kirby
Mark A
MA
Sussman
Gilles
G
Lajoie
Jennifer E
JE
Van Eyk
D000818:Animals; D002311:Cardiomyopathy, Dilated / Q000201:enzymology* / Q000378:metabolism; D003402:Creatine Kinase / Q000032:analysis* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D006333:Heart Failure / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism; D018994:Myosin Light Chains / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020830:rac1 GTP-Binding Protein / Q000032:analysis* / Q000378:metabolism
D018994:Myosin Light Chains; D003402:Creatine Kinase; D020830:rac1 GTP-Binding Protein