title: Phosphorylation of dystrophin. The carboxyl-terminal region of dystrophin is a substrate for in vitro phosphorylation by p34cdc2 protein kinase.
abstract: In this paper, we report that p34cdc2 protein kinase phosphorylates recombinant fragments of skeletal muscle dystrophin with a maximal incorporation of 1.8 mol of Pi/mol of protein. Phosphorylation of both serine and threonine residues occurs within the carboxyl-terminal 201 amino acids of dystrophin, with phosphothreonine localized to within 25 residues of the carboxyl terminus. Supporting these in vitro studies, we also show that native dystrophin is phosphorylated by p34cdc2 kinase in isolated sarcolemmal vesicles. Sequence analysis indicates two consensus sites for p34cdc2 protein kinase within the carboxyl-terminal 201 amino acids of dystrophin. Importantly, neither of these sites is conserved in dystrophin-related protein, and only one site is conserved in the 71-kDa alternative product of the Duchenne muscular dystrophy gene, despite an otherwise extremely high degree of sequence conservation between these proteins. Importantly, in this study we also show that dystrophin is phosphorylated in vivo in rat skeletal muscle primary cultures, and we suggest that further investigation of both in vivo and in vitro phosphorylation of this protein will comprise an important part in determination of its function(s).
authors.affiliation: Cardiovascular Disease Research Group, University of Alberta, Edmonton, Canada.
authors.forename: R E
authors.initials: RE
authors.lastname: Milner
authors.forename: J L
authors.initials: JL
authors.lastname: Busaan
authors.forename: C F
authors.initials: CF
authors.lastname: Holmes
authors.forename: J H
authors.initials: JH
authors.lastname: Wang
authors.forename: M
authors.initials: M
authors.lastname: Michalak
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002478:Cells, Cultured; D016189:Dystrophin / Q000378:metabolism*; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D012508:Sarcolemma / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D016189:Dystrophin; D016203:CDC2 Protein Kinase