Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research


Carberry, Steven and Zweyer, Margit and Swandulla, Dieter and Ohlendieck, Kay (2013) Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research. Biology, 2. pp. 1438-1464. ISSN 2079-7737

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Abstract

In this article, we illustrate the application of difference in-gel electrophoresis for the proteomic analysis of dystrophic skeletal muscle. The mdx diaphragm was used as a tissue model of dystrophinopathy. Two-dimensional gel electrophoresis is a widely employed protein separation method in proteomic investigations. Although two-dimensional gels usually underestimate the cellular presence of very high molecular mass proteins, integral membrane proteins and low copy number proteins, this method is extremely powerful in the comprehensive analysis of contractile proteins, metabolic enzymes, structural proteins and molecular chaperones. This gives rise to two-dimensional gel electrophoretic separation as the method of choice for studying contractile tissues in health and disease. For comparative studies, fluorescence difference in-gel electrophoresis has been shown to provide an excellent biomarker discovery tool. Since aged diaphragm fibres from the mdx mouse model of Duchenne muscular dystrophy closely resemble the human pathology, we have carried out a mass spectrometry-based comparison of the naturally aged diaphragm versus the senescent dystrophic diaphragm. The proteomic comparison of wild type versus mdx diaphragm resulted in the identification of 84 altered protein species. Novel molecular insights into dystrophic changes suggest increased cellular stress, impaired calcium buffering, cytostructural alterations and disturbances of mitochondrial metabolism in dystrophin-deficient muscle tissue.*

Item Type: Article
Additional Information: This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited: Carberry, S.; Zweyer, M.; Swandulla, D.; Ohlendieck, K. Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research. Biology 2013, 2, 1438-1464.
Keywords: cvHsp; diaphragm; DIGE; Duchenne muscular dystrophy; dystrophinopathy; gel electrophoresis; HspB7; mdx; mouse model; parvalbumin;
Academic Unit: Faculty of Science and Engineering > Biology
Item ID: 6900
Identification Number: 10.3390/biology2041438
Depositing User: Prof. Kay Ohlendieck
Date Deposited: 21 Jan 2016 14:53
Journal or Publication Title: Biology
Publisher: MDPI
Refereed: Yes
Funders: Muscular Dystrophy Ireland, Duchenne Ireland, Hume Scholarship, Health Research Board (HRB), Higher Education Authority (HEA)
URI:

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