title: Proteinase-sensitive sites on isolated rabbit dystrophin.
abstract: Dystrophin was isolated from the purified large oligomeric dystrophin complex with its associated proteins (DC) of rabbit skeletal muscle by alkaline dissociation followed by gel filtration to remove the associated proteins. Isolated dystrophin and DC were subjected to digestion with calpain or alpha-chymotrypsin, and the generated polypeptide fragments were studied by immunoblot analysis using seven kinds of antibodies raised against antigens corresponding to various regions from the N- to the C-terminal of human dystrophin. For some fragments, the amino acid sequences at the N-termini were determined. Two proteinases, which bear distinct specificities, generated very similar fragments from purified dystrophin with or without the associated proteins. The cleavage sites found by mapping the fragments onto the dystrophin molecule were similar to those found in a previous study using crude mouse muscle cell membrane fraction [Koenig, M. & Kunkel, L.M. (1990) J. Biol. Chem. 265, 4560-4566]. On the basis of these results, we concluded that dystrophin has several unique proteinase-sensitive sites.
authors: Division of Cell Biology, National Institute of Neuroscience, NCNP, Tokyo.
authors: M
authors: M
authors: Yoshida
authors: A
authors: A
authors: Suzuki
authors: T
authors: T
authors: Shimizu
authors: E
authors: E
authors: Ozawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002154:Calpain / Q000378:metabolism / Q000494:pharmacology; D002850:Chromatography, Gel; D002918:Chymotrypsin / Q000378:metabolism / Q000494:pharmacology; D016189:Dystrophin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D010449:Peptide Mapping; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D016189:Dystrophin; D010446:Peptide Fragments; D010450:Endopeptidases; D002918:Chymotrypsin; D002154:Calpain