Transfer of polymorphic monoclonal antibody epitopes to the first and second domains of HLA-DR beta-chains by site-directed mutagenesis.
We used site-directed mutagenesis of HLA-DR beta-chains to localize the binding sites for two polymorphic DR-binding mAb to residues in the first and second external domains, respectively. Transfer of three first domain alpha-helical residues, G73, R74 and N77, normally present in DR3a and DRw52a, to a DR4 beta-chain was sufficient for recognition of this mutant DR molecule by a DR3-specific mAb, NDS 9. A polymorphism controlling the binding of a DR4-specific mAb, GS 359-13F10, was mapped to a tyrosine at position 96 of the DR4 beta-chain second domain by the construction fo a chimeric DR molecule containing a DR2-first domain and DR4-second domain. The mapping of these two polymorphic epitopes to specific positions on the DR beta-chain will allow further structural and functional analysis of the DR molecule.
Maurer
D
D
Immunogenetics Research Section, Blood Center of Southeastern Wisconsin, Milwaukee 53233.
Gorski
J
J
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D006684:HLA-DR Antigens / Q000235:genetics* / Q000276:immunology*; D015802:HLA-DR2 Antigen / Q000235:genetics / Q000276:immunology; D015803:HLA-DR3 Antigen / Q000235:genetics / Q000276:immunology; D015804:HLA-DR4 Antigen / Q000235:genetics / Q000276:immunology; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011110:Polymorphism, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006684:HLA-DR Antigens; D015802:HLA-DR2 Antigen; D015803:HLA-DR3 Antigen; D015804:HLA-DR4 Antigen; D011993:Recombinant Fusion Proteins