title: Computer analysis of mutations that affect antibody specificity.
abstract: The mouse hybridoma cell line 40-150 secretes antibodies with high affinity toward the cardiac glycosides digoxin and digitoxin. A spontaneous mutant, 40-150 A2.4, produces an antibody which carries a single residue mutation, Ser----Arg, in its heavy chain (H94) and has an altered specificity. A second-order mutant, 40-150 A2.4 P.10, produces two antibody molecules, one the same as 40-150 A2.4, the other lacking two residues at the N-terminus of its H chain, and having a specificity profile approaching that of 40-150 antibody. The N-terminus and the position H94 are distant from the antigen-binding site of the antibody; thus, the structural basis of the specificity changes was not immediately clear. Approximate structures of the 40-150 antibody and its mutants were constructed in the computer, based on atomic coordinates of the homologous mouse antibody McPC 603. Using the program CONGEN, the torsional space of the polypeptide backbone and side chains around position H94 was uniformly sampled, and the lowest energy conformations were analyzed in detail. The results indicate that when Arg-H94 is substituted for Ser, Arg-H94 can hydrogen bond to side chains of Asp-H101, Arg-L46, and Asp-L55. This results in a change in the surface of the combining site which may account for the affinity changes. Deletion of the two N-terminal residues increases solvent accessibility of Arg-H94. The solvation may cause a hydrogen bond between Arg-H94 and Asp-H101 to be lost, restoring the structure to one similar to that of 40-150.
authors.affiliation: Massachusetts General Hospital, Boston 02114.
authors.forename: J
authors.initials: J
authors.lastname: Novotny
authors.forename: R E
authors.initials: RE
authors.lastname: Bruccoleri
authors.forename: E
authors.initials: E
authors.lastname: Haber
mesh_terms: D000818:Animals; D000918:Antibody Specificity / Q000235:genetics*; D002460:Cell Line; D001330:Electronic Data Processing; D006801:Humans; D006825:Hybridomas; D006860:Hydrogen Bonding; D007136:Immunoglobulins / Q000235:genetics; D051379:Mice; D009154:Mutation; D011487:Protein Conformation
chemical_list: D007136:Immunoglobulins