Epitope mapping of antibodies to acetylcholine receptor alpha subunits using peptides synthesized on polypropylene pegs.
Concurrent synthesis of overlapping octameric peptides corresponding to the sequence of the Torpedo acetylcholine receptor (AChR) alpha subunit has been carried out on polypropylene supports functionalized with primary amino groups according to a method developed by M. Geysen [(1987) J. Immunol. Methods 102, 259-274]. The peptides on the solid supports have been used in an enzyme-linked immunosorbent assay. Interactions of the synthetic peptides with antibodies are then detected without removing them from the solid support. By this procedure, epitopes of both antisera and monoclonal antibodies to the Torpedo acetylcholine receptor, its subunits, and synthetic peptide fragments have been mapped. Both rat and rabbit antisera to the alpha subunit show major epitopes spanning the residues 150-165, 338-345, and 355-366 on the Torpedo AChR alpha subunit. Epitopes of monoclonal antibodies to these major epitopes and to others have been rather precisely mapped by using this technique with peptides of varying lengths. The specificity of several of these mAbs are of interest because they have been used in mapping the transmembrane orientation of the AChR alpha-subunit polypeptide chain.
Das
M K
MK
Receptor Biology Laboratory, Salk Institute for Biological Studies, San Diego, California 92138.
Lindstrom
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000939:Epitopes / Q000032:analysis*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis*; D011126:Polypropylenes; D011487:Protein Conformation; D011950:Receptors, Cholinergic / Q000276:immunology*; D014101:Torpedo
D000911:Antibodies, Monoclonal; D000939:Epitopes; D046911:Macromolecular Substances; D009842:Oligopeptides; D011126:Polypropylenes; D011950:Receptors, Cholinergic