Doran, Philip and Dowling, Paul and Lohan, James and McDonnell, Karen and Poetsch, Stephan and Ohlendieck, Kay (2004) Subproteomics analysis of Ca2+-binding proteins demonstrates decreased calsequestrin expression in dystrophic mouse skeletal muscle. European Journal of Biochemistry, 271 (19). pp. 3943-3952. ISSN 0014-2956

Preview

Download (311kB) | Preview

Abstract

Duchenne muscular dystrophy represents one of the most common hereditary diseases. Abnormal ion handling is believed to render dystrophin-deficient muscle fibres more susceptible to necrosis. Although a reduced Ca(2+) buffering capacity has been shown to exist in the dystrophic sarcoplasmic reticulum, surprisingly no changes in the abundance of the main luminal Ca(2+) reservoir protein calsequestrin have been observed in microsomal preparations. To address this unexpected finding and eliminate potential technical artefacts of subcellular fractionation protocols, we employed a comparative subproteomics approach with total mouse skeletal muscle extracts. Immunoblotting, mass spectrometry and labelling of the entire muscle protein complement with the cationic carbocyanine dye 'Stains-All' was performed in order to evaluate the fate of major Ca(2+)-binding proteins in dystrophin-deficient skeletal muscle fibres. In contrast to a relatively comparable expression pattern of the main protein population in normal vs. dystrophic fibres, our analysis showed that the expression of key Ca(2+)-binding proteins of the luminal sarcoplasmic reticulum is drastically reduced. This included the main terminal cisternae constituent, calsequestrin, and the previously implicated Ca(2+)-shuttle element, sarcalumenin. In contrast, the 'Stains-All'-positive protein spot, representing the cytosolic Ca(2+)-binding component, calmodulin, was not changed in dystrophin-deficient fibres. The reduced 2D 'Stains-All' pattern of luminal Ca(2+)-binding proteins in mdx preparations supports the calcium hypothesis of muscular dystrophy. The previously described impaired Ca(2+) buffering capacity of the dystrophic sarcoplasmic reticulum is probably caused by a reduction in luminal Ca(2+)-binding proteins, including calsequestrin.

Item Type:	Article
Keywords:	calsequestrin; mdx; mouse skeletal muscle; muscular dystrophy; sarcalumenin;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7358
Identification Number:	https://doi.org/10.1111/j.1432-1033.2004.04332.x
Depositing User:	Paul Dowling
Date Deposited:	18 Aug 2016 15:22
Journal or Publication Title:	European Journal of Biochemistry
Publisher:	Wiley-Blackwell
Refereed:	Yes
Funders:	European Commission, Muscular Dystrophy Ireland, Health Research Board (HRB)
URI:	http://onlinelibrary.wiley.com
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

Repository Staff Only(login required)

Item control page

Downloads

Origin of downloads