title: The molecular basis for cross-reaction of an anti-dystrophin antibody with alpha-actinin.
abstract: The epitope recognised by the anti-dystrophin monoclonal antibodies MANDYS141 and MANDYS142 has been characterised using a phage display peptide library and a bacteriophage lambda cDNA library. Using a phage display library of random 15-mer peptides, the epitope recognised by the two antibodies was identified as EEXF. A lambda gt11 clone obtained by screening a human muscle cDNA library was shown to contain part of the out-of-frame human mitochondrial succinyl CoA synthetase (alpha-subunit) cDNA sequence which contains the sequence EEPL, suggesting a minimum requirement of EEXF/L for antibody binding. The sequence EEDF is located in the helical rod region of dystrophin and the N-terminal domain of alpha-actinin; this may explain why native dystrophin is not detected, since the alpha-helical, coiled-coil folding of the rod region of dystrophin may obscure the epitope in the native protein. The antibody cross-reaction between dystrophin and alpha-actinin is likely to be fortuitous and not due to any structural homology that exists between these two members of the spectrin superfamily.
authors.affiliation: MRIC Biochemistry Group, The North East Wales Institute, Wrexham, Clwyd, UK.
authors.forename: M
authors.initials: M
authors.lastname: James
authors.forename: N T
authors.initials: NT
authors.lastname: Man
authors.forename: Y H
authors.initials: YH
authors.lastname: Edwards
authors.forename: G E
authors.initials: GE
authors.lastname: Morris
mesh_terms: D000185:Actinin / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003429:Cross Reactions; D016189:Dystrophin / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D008969:Molecular Sequence Data; D051381:Rats; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D016189:Dystrophin; D000185:Actinin