Purification and characterization of the 1, 200-kDa subfragment of connectin filaments produced by 0.1 mM calcium ions

Ryoichi Tanabe, Ryuichi Tatsumi, Koui Takahashi

研究成果: ジャーナルへの寄稿学術誌査読

27 被引用数 (Scopus)

抄録

When myofibrils prepared from chicken leg muscle were treated with a solution containing 0.1 mM CaCl2 and 30 μg/ml of leupeptin, α-connectin, which exists as a longitudinal thin filament in a sarcomere, was split into β-connectin and α 1, 200-kDa subfragment. The native subfragment was successfully purified without using any denaturant: It was extracted with 1 M KI solution from the Ca-treated myofibrils and purified by TSKge1 G6000PW column chromatography. About 10 mg of the subfragment was yielded from 100 g of starting muscle. Using immunofluorescence microscopy and immunoelectron microscopy, we show here that polyclonal antibodies against the 1, 200-kDa subfragment bind to the Z-disk and the epitope, which is about 0.34 um apart from the Z-disk at a sarcomere length of 2.6 μm; the 1, 200-kDa subfragment constitutes the proximal region of connectin filaments. Purified α-actinin decorated α-connectin and the 1, 200-kDa subfragment on nitrocellulose blots of myofibrillar proteins separated by SDS-PAGE. Therefore, we conclude that connectin filaments are anchored to the Z-disk by the binding of the 1, 200-kDa subfragment to α-actinin.

本文言語英語
ページ(範囲)351-355
ページ数5
ジャーナルJournal of biochemistry
115
2
DOI
出版ステータス出版済み - 1月 1 1994
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学

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