Two-dimensional gel electrophoresis of actin-binding proteins isolated by affinity chromatography from human skeletal muscle

Title
Two-dimensional gel electrophoresis of actin-binding proteins isolated by affinity chromatography from human skeletal muscle
Publication Date
1997
Author(s)
Coumans-Moens, Joelle
( author )
OrcID: https://orcid.org/0000-0001-6642-5202
Email: jmoensco@une.edu.au
UNE Id une-id:jmoensco
Humphery-Smith, Ian
dos Remedios, Cristobal G
Type of document
Journal Article
Language
en
Entity Type
Publication
Publisher
Wiley-VCH Verlag GmbH & Co KGaA
Place of publication
Germany
DOI
10.1002/elps.1150180709
UNE publication id
une:23114
Abstract
In muscle cells actin exists as a mixture of monomeric (G-actin) and filamentous actin (F-actin) and ionic conditions strongly favor the formation of F-actin. The existence of unpolymerized actin depends, among other factors, on proteins that bind to G-actin, the so-called G-actin-binding proteins (G-ABPs). We have coupled monomeric actin to divinylsulphone-activated agarose (Mini-Leak) to isolate G-ABPs in human skeletal muscle. Eluted proteins were analyzed by two‐dimensional gel electrophoresis (2-DE), which shows that some proteins are selectively retained. Deoxyribonuclease I (DNase I) is known to bind residues at the "pointed end" of actin (subdomains 2 and 4) with a high affinity. When DNase I is bound to the actin Mini‐Leak before applying the skeletal muscle extract, the 2-DE gels of the eluted proteins reveals differences when compared to gels of proteins eluted from actin-Mini-Leak and DNase I-Mini‐Leak affinity columns. This strategy should detect ABPs which bind to sites other than the DNase I-binding site and some may prove to be novel.
Link
Citation
Electrophoresis, 18(7), p. 1079-1085
ISSN
1522-2683
0173-0835
Start page
1079
End page
1085

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