title: Disruption of the utrophin-actin interaction by monoclonal antibodies and prediction of an actin-binding surface of utrophin.
abstract: Monoclonal antibody (mAb) binding sites in the N-terminal actin-binding domain of utrophin have been identified using phage-displayed peptide libraries, and the mAbs have been used to probe functional regions of utrophin involved in actin binding. mAbs were characterized for their ability to interact with the utrophin actin-binding domain and to affect actin binding to utrophin in sedimentation assays. One of these antibodies was able to inhibit utrophin-F-actin binding and was shown to recognize a predicted helical region at residues 13-22 of utrophin, close to a previously predicted actin-binding site. Two other mAbs which did not affect actin binding recognized predicted loops in the second calponin homology domain of the utrophin actin-binding domain. Using the known three-dimensional structure of the homologous actin-binding domain of fimbrin, these results have enabled us to determine the likely orientation of the utrophin actin-binding domain with respect to the actin filament.
authors.affiliation: MRIC Biochemistry Group, N.E. Wales Institute, Mold Road, Wrexham LL1 2AW, Wales, U.K.
authors.forename: G E
authors.initials: GE
authors.lastname: Morris
authors.forename: T M
authors.initials: TM
authors.lastname: Nguyen
authors.forename: T N
authors.initials: TN
authors.lastname: Nguyen
authors.forename: A
authors.initials: A
authors.lastname: Pereboev
authors.forename: J
authors.initials: J
authors.lastname: Kendrick-Jones
authors.forename: S J
authors.initials: SJ
authors.lastname: Winder
mesh_terms: D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism*; D018604:Epitope Mapping; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011817:Rabbits; D049411:Utrophin
chemical_list: D000199:Actins; D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D049411:Utrophin