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Titolo:
PURIFICATION AND CHARACTERIZATION OF THE 1,200-KDA SUBFRAGMENT OF CONNECTIN FILAMENTS PRODUCED BY 0.1 MM CALCIUM-IONS
Autore:
TANABE R; TATSUMI R; TAKAHASHI K;
Indirizzi:
HOKKAIDO UNIV,FAC AGR,DEPT ANIM SCI,MEAT SCI LAB,KITA KU SAPPORO HOKKAIDO 060 JAPAN HOKKAIDO UNIV,FAC AGR,DEPT ANIM SCI,MEAT SCI LAB,KITA KU SAPPORO HOKKAIDO 060 JAPAN
Titolo Testata:
Journal of Biochemistry
fascicolo: 2, volume: 115, anno: 1994,
pagine: 351 - 355
SICI:
0021-924X(1994)115:2<351:PACOT1>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
FROG SKELETAL-MUSCLE; IMMUNOELECTRON MICROSCOPY; ALPHA-CONNECTIN; MONOCLONAL-ANTIBODIES; ELASTIC PROTEIN; TITIN; NEBULIN; ACTININ; EPITOPES; HEAD;
Keywords:
BINDING OF ALPHA-ACTININ; LOCALIZATION OF 1,200-KDA SUBFRAGMENT; PURIFICATION OF 1,200-KDA SUBFRAGMENT; 1,200-KDA SUBFRAGMENT OF CONNECTIN FILAMENTS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
28
Recensione:
Indirizzi per estratti:
Citazione:
R. Tanabe et al., "PURIFICATION AND CHARACTERIZATION OF THE 1,200-KDA SUBFRAGMENT OF CONNECTIN FILAMENTS PRODUCED BY 0.1 MM CALCIUM-IONS", Journal of Biochemistry, 115(2), 1994, pp. 351-355

Abstract

When myofibrils prepared from chicken leg muscle were treated with a solution containing 0.1 mM CaCl2 and 30 mu g/ml of leupeptin, alpha-connectin, which exists as a longitudinal thin filament in a sarcomere, was split into beta-connectin and a 1,200-kDa subfragment. The native subfragment was successfully purified without using any denaturant: Itwas extracted with 1M KI solution from the Ca-treated myofibrils and purified by TSKgel 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 mu m apart from the Z-disk ata sarcomere length of 2.6 mu m; the 1,200-kDa subfragment constitutesthe proximal region of connectin filaments. Purified alpha-actinin decorated alpha-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 alpha-actinin.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/07/20 alle ore 22:21:14