Please use this identifier to cite or link to this item: https://hdl.handle.net/1959.11/22926
Title: Actin-binding proteins in mouse C2 myoblasts and myotubes: A combination of affinity chromatography and two-dimensional gel electrophoresis
Contributor(s): Coumans-Moens, Joelle (author)orcid ; dos Remedios, Cristobal G (author)
Publication Date: 1998
DOI: 10.1002/elps.1150190537
Handle Link: https://hdl.handle.net/1959.11/22926
Abstract: This paper analyzes proteins expressed in a mouse muscle precursor cell line (C2 myoblasts) and compares them with those observed in differentiated myotubes from the same cell line. We observed hundreds of proteins in myoblasts using IPG two‐dimensional gel electrophoresis but this number is greatly reduced using Mini‐Leak (divinylsulfone‐activated agarose) affinity chromatography. Two kinds of affinity columns were prepared. One contained a chemically modified monomeric actin bound to the affinity matrix. The second matrix contained a high‐affinity actin‐binding protein (DNase I) which was bound to the actin Mini‐Leak column to block specific sites on actin. Actin‐binding proteins in homogenates of myoblasts or myotubes were passed through the affinity columns and eluted under high salt conditions. The Mini‐Leak affinity medium itself appeared to have little ability to bind proteins. Our two‐dimensional (2‐D) gels identified a small number of proteins and we are currently focusing our attention on a particular protein spot which could correspond to cofilin. Comparison of myoblast and myotube proteins using affinity chromatography shows no qualitative, clearly identifiable differences but the analysis is still in progress. These findings are discussed in relation to reports in which the myoblast‐myotube transformation was associated with the up‐regulation or de novo synthesis of more than ten proteins.
Publication Type: Journal Article
Source of Publication: Electrophoresis, 19(5), p. 826-833
Publisher: Wiley-VCH Verlag GmbH & Co KGaA
Place of Publication: Germany
ISSN: 1522-2683
0173-0835
Field of Research (FOR): 060101 Analytical Biochemistry
060103 Cell Development, Proliferation and Death
060109 Proteomics and Intermolecular Interactions (excl. Medical Proteomics)
Peer Reviewed: Yes
HERDC Category Description: C1 Refereed Article in a Scholarly Journal
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