Mutational analysis of the fine specificity of binding of monoclonal antibody 51F to lambda repressor.
Monoclonal antibody 51F recognizes determinants in the helix 4 region of the native form of the N-terminal domain of lambda repressor. A cassette mutagenesis method was used to introduce changes within this region, and antibody-reactive candidates were isolated and sequenced. The resulting data allow the identification of repressor side chains that are critical determinants of antibody binding. Four of these side chains are on the surface of the N-terminal domain and probably contact the antibody directly. These contact positions were then mutagenized individually, and the antibody binding phenotypes of a large number of singly mutant repressors were determined. Taken together, the mutational data allow a functional map of the recognition surface to be constructed and the physical nature of some of the specific interactions that stabilize the antibody-antigen complex to be surmised.
Breyer
R M
RM
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Sauer
R T
RT
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000235:genetics*; D000914:Antibodies, Viral / Q000032:analysis / Q000235:genetics; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012097:Repressor Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics*; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins