title: Isolation of nebulin from rabbit skeletal muscle and its interaction with actin.
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.
authors.affiliation: Department of Biology, Graduate School of Science, Chiba University, Chiba 263-8522, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Chitose
authors.forename: Atsushi
authors.initials: A
authors.lastname: Watanabe
authors.forename: Masato
authors.initials: M
authors.lastname: Asano
authors.forename: Akira
authors.initials: A
authors.lastname: Hanashima
authors.forename: Kouhei
authors.initials: K
authors.lastname: Sasano
authors.forename: Yulong
authors.initials: Y
authors.lastname: Bao
authors.forename: Koscak
authors.initials: K
authors.lastname: Maruyama
authors.forename: Sumiko
authors.initials: S
authors.lastname: Kimura
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D000837:Anion Exchange Resins; D015153:Blotting, Western; D002852:Chromatography, Ion Exchange / Q000379:methods*; D009124:Muscle Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry*; D011817:Rabbits; D014335:Tropomyosin; D014336:Troponin; D014508:Urea / Q000737:chemistry
chemical_list: D000199:Actins; D000837:Anion Exchange Resins; D009124:Muscle Proteins; D014335:Tropomyosin; D014336:Troponin; C035620:nebulin; D014508:Urea
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