Identification of HLA-DR1 beta chain residues critical for binding staphylococcal enterotoxins A and E.
Superantigens are thought to make external contacts with major histocompatibility complex (MHC) class II molecules and with the V beta portion of a T cell antigen receptor (TCR), thereby stimulating entire families of T cells. The precise mapping of superantigen binding sites on class II molecules may provide valuable information on how TCR and MHC molecules interact. Two bacterial superantigens, staphylococcal enterotoxins A and E (SEA/SEE) bind well to most HLA-DR alleles, but poorly to HLA-DRw53. The sequences responsible for this binding were localized to the putative alpha helix of the DR beta chain by measuring toxin binding to a panel of chimeric class II molecules expressed on transfected cells. Binding of SEA/SEE to the DRw14 (Dw9) molecule suggested that the conserved histidine 81 in the beta chain of most DR molecules was important, whereas the tyrosine 81 in the DRw53 beta chain was detrimental for high-affinity binding. To prove this, reciprocal point mutations were introduced in the DR1 and DRw53 beta chains. Mutation of histidine 81 in the DR1 beta chain to tyrosine reduced SEA/SEE binding, but did not prevent recognition of two DR1-restricted peptides by six of eight antigen-specific T cell lines. Conversely, introduction to histidine at position 81 in the DRw53 beta chain restored normal levels of SEA/SEE binding. These data suggest that a binding site of SEA and SEE lies on the outer face of the beta chain alpha helix, pointing away from the antigen-binding groove.
Karp
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Simmons Arthritis Research Center, University of Texas Southwestern Medical Center, Dallas 75235-8884.
Long
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D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004269:DNA, Bacterial / Q000378:metabolism; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes / Q000378:metabolism*; D006684:HLA-DR Antigens / Q000235:genetics / Q000378:metabolism; D059810:HLA-DR Serological Subtypes; D015798:HLA-DR1 Antigen / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D059845:HLA-DRB4 Chains; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D013211:Staphylococcus aureus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection
D004269:DNA, Bacterial; D004768:Enterotoxins; D000939:Epitopes; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; D015798:HLA-DR1 Antigen; C052238:HLA-DR14; C050610:HLA-DR53; D059845:HLA-DRB4 Chains; D016693:Receptors, Antigen, T-Cell, alpha-beta; C049463:enterotoxin E, Staphylococcal; C026066:enterotoxin A, Staphylococcal