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Journal of Biomedicine and Biotechnology
Volume 2010, Article ID 108495, 8 pages
Research Article

Isolation of Nebulin from Rabbit Skeletal Muscle and Its Interaction with Actin

Department of Biology, Graduate School of Science, Chiba University, Chiba 263-8522, Japan

Received 1 December 2009; Accepted 15 February 2010

Academic Editor: Henk L. M. Granzier

Copyright © 2010 Ryo Chitose et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Nebulin is about 800 kDa filamentous protein that binds the entire thin filament of vertebrate skeletal muscle sarcomeres. Nebulin cannot be isolated from muscle except in a completely denatured form by direct solubilization of myofibrils with SDS because nebulin is hardly soluble under salt conditions. In the present study, nebulin was solubilized by a salt solution containing 1 M urea and purified by DEAE-Toyopearl column chromatography via 4 M urea elution. Rotary-shadowed images of nebulin showed entangled knit-like particles, about 20 nm in diameter. The purified nebulin bound to actin filaments to form loose bundles. Nebulin was confirmed to bind actin, -actinin, -actinin, and tropomodulin, but not troponin or tropomyosin. The data shows that full-length nebulin can be also obtained in a functional and presumably native form, verified by data from experiments using recombinant subfragments.