Subproteomic analysis of basic proteins in aged skeletal muscle following offgel pre-fractionation

  • Authors:
    • Joan Gannon
    • Kay Ohlendieck
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  • Published online on: January 17, 2012     https://doi.org/10.3892/mmr.2012.759
  • Pages: 993-1000
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Abstract

The progressive loss of skeletal muscle mass is a serious pathophysiological problem in the elderly, which warrants detailed biochemical studies into the underlying mechanism of age-related fiber degeneration. Over the last few years, mass spectrometry (MS)-based proteomics has identified a considerable number of new biomarkers of muscle aging in humans and animal models of sarcopenia. However, interpretation of the proteomic findings is often complicated by technical and biological limitations. Although gel electrophoresis-based approaches represent a highly sensitive analytical way for the large-scale and high-throughput survey of global changes in skeletal muscle proteins during aging, often the presence of components with an isoelectric point in the basic range is underestimated. We, therefore, carried out a comparative subproteomic study of young versus aged rat muscle focusing on potential changes in muscle proteins with an alkaline isoelectric point, using a combination of offgel electrophoresis and two-dimensional (2D) slab gel electrophoresis. Offgel electrophoresis was successfully applied as a prefractionation step to enrich basic protein species from crude tissue extracts representing young adult versus senescent muscle specimens. Proteomics has demonstrated alterations in a small cohort of basic proteins during muscle aging. The mass spectrometric identification of altered proteins and immunoblotting revealed a decrease in the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and a concomitant increase in mitochondrial creatine kinase (CK) and ubiquinol cytochrome‑c reductase. This agrees with the idea of a glycolytic-to-oxidative shift during muscle aging, which is indicative of an overall fast-to-slow transition process in senescent rat muscle. Thus, alterations in the abundance of metabolic enzymes appear to play a central role in the molecular pathogenesis of age‑dependent muscle wasting.
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April 2012
Volume 5 Issue 4

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Gannon J and Gannon J: Subproteomic analysis of basic proteins in aged skeletal muscle following offgel pre-fractionation. Mol Med Rep 5: 993-1000, 2012
APA
Gannon, J., & Gannon, J. (2012). Subproteomic analysis of basic proteins in aged skeletal muscle following offgel pre-fractionation. Molecular Medicine Reports, 5, 993-1000. https://doi.org/10.3892/mmr.2012.759
MLA
Gannon, J., Ohlendieck, K."Subproteomic analysis of basic proteins in aged skeletal muscle following offgel pre-fractionation". Molecular Medicine Reports 5.4 (2012): 993-1000.
Chicago
Gannon, J., Ohlendieck, K."Subproteomic analysis of basic proteins in aged skeletal muscle following offgel pre-fractionation". Molecular Medicine Reports 5, no. 4 (2012): 993-1000. https://doi.org/10.3892/mmr.2012.759