title: Mapping epitopes on the (Ca(2+)-Mg2+)-ATPase of sarcoplasmic reticulum using fusion proteins.
abstract: Epitopes for a number of monoclonal antibodies (mAbs) binding (Ca(2+)-Mg2+)-ATPase purified from skeletal muscle sarcoplasmic reticulum have been defined by studying binding to fusion proteins generated from cDNA fragment libraries. Comparison of these results with those of previous studies of binding of mAbs to proteolytic fragments of the ATPase have allowed the definition of the epitopes to within approx. 100 residues and for one (mAb 1/2H7) to within 45 residues. The experiments suggest considerable exposure of the nucleotide binding domain of the ATPase on the top surface of the protein. Those mAbs that were found to inhibit steady-state ATPase activity were found to bind to epitopes in the nucleotide binding domain of the ATPase.
authors.affiliation: Department of Biochemistry, University of Southampton, U.K.
authors.forename: R E
authors.initials: RE
authors.lastname: Tunwell
authors.forename: C D
authors.initials: CD
authors.lastname: O'Connor
authors.forename: A M
authors.initials: AM
authors.lastname: Mata
authors.forename: J M
authors.initials: JM
authors.lastname: East
authors.forename: A G
authors.initials: AG
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D017301:Ca(2+) Mg(2+)-ATPase / Q000276:immunology*; D000252:Calcium-Transporting ATPases / Q000276:immunology*; D000939:Epitopes / Q000032:analysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases