Carberry, Steven and Zweyer, Margit and Swandulla, Dieter and Ohlendieck, Kay
(2014)
Comparative proteomic analysis of the contractile-protein-depleted
fraction from normal versus dystrophic skeletal muscle.
Analytical Biochemistry, 446.
pp. 108-115.
ISSN 0003-2697
Abstract
In basic and applied myology, gel-based proteomics is routinely used for studying global changes in the
protein constellation of contractile fibers during myogenesis, physiological adaptations, neuromuscular
degeneration, and the natural aging process. Since the main proteins of the actomyosin apparatus and
its auxiliary sarcomeric components often negate weak signals from minor muscle proteins during proteomic
investigations, we have here evaluated whether a simple prefractionation step can be employed
to eliminate certain aspects of this analytical obstacle. To remove a large portion of highly abundant contractile
proteins from skeletal muscle homogenates without the usage of major manipulative steps, differential
centrifugation was used to decisively reduce the sample complexity of crude muscle tissue
extracts. The resulting protein fraction was separated by two-dimensional gel electrophoresis, and 2Dlandmark
proteins were identified by mass spectrometry. To evaluate the suitability of the contractileprotein-
depleted fraction for comparative proteomics, normal versus dystrophic muscle preparations
were examined. The mass spectrometric analysis of differentially expressed proteins, as determined by
fluorescence difference in-gel electrophoresis, identified 10 protein species in dystrophic mdx hindlimb
muscles. Interesting new biomarker candidates included Hsp70, transferrin, and ferritin, whereby their
altered concentration levels in dystrophin-deficient muscle were confirmed by immunoblotting.
| Item Type: |
Article
|
| Keywords: |
Dystrophinopathy; Ferritin; Hsp70; Muscle proteomics; Transferrin; |
| Academic Unit: |
Faculty of Science and Engineering > Biology |
| Item ID: |
7487 |
| Identification Number: |
https://doi.org/10.1016/j.ab.2013.08.004 |
| Depositing User: |
Prof. Kay Ohlendieck
|
| Date Deposited: |
07 Oct 2016 10:44 |
| Journal or Publication Title: |
Analytical Biochemistry |
| Publisher: |
Elsevier |
| Refereed: |
Yes |
| Funders: |
Muscular Dystrophy Ireland, Duchenne Ireland, Health Research Board (HRB) |
| URI: |
|
| Use Licence: |
This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available
here |
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