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

Overview

Journal J Biomed Biotechnol

Specialty Biology

Date 2010 May 15

PMID 20467585

Citations 10

Authors

Ryo Chitose

Atsushi Watanabe

Masato Asano

Akira Hanashima

Kouhei Sasano

Yulong Bao

Koscak Maruyama

Sumiko Kimura

Affiliations

Soon will be listed here.

Abstract

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, alpha-actinin, beta-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.

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