Localization of binding sites of staphylococcal enterotoxin B (SEB), a superantigen, for HLA-DR by inhibition with synthetic peptides of SEB.
Staphylococcal enterotoxins are major causes of food poisoning and toxic shock syndrome. Their ability to bind to major histocompatibility complex (MHC) class II molecules has been suggested to be the first step in the mechanism whereby they cause illness. By flow cytometric analysis, the sites of interaction of staphylococcal enterotoxin B (SEB) with HLA-DR molecules were probed in the present study by inhibiting the binding of biotinylated SEB to a human T-cell line (HUT-78) with synthetic peptides of SEB. Five peptides of SEB gave significant inhibition of binding: a peptide containing amino acids 9 to 20 [SEB(9-20)], SEB(30-38), SEB(61-70), SEB(90-114), and SEB(169-181). One peptide, SEB(39-51), enhanced binding. Among the inhibitory peptides, SEB(90-114), a peptide spanning the entire disulfide loop, showed the most efficient inhibition of binding. Peptides SEB(9-20) and SEB(39-51) include amino acid residues that have been identified by previous mutation studies (J.W. Kappler, A. Herman, J. Clements, and P. Marrack, J. Exp. Med. 175:387-396, 1992) as being important in binding to MHC class II. Amino acids lining the alpha 5 groove of SEB have also been postulated to be involved in binding to MHC class II molecules. However, only two of the residues that line the alpha 5 groove of SEB, His-12 and Tyr-17, are on peptide SEB(9-20) that inhibits binding. These results confirm previous studies that implicated the amino-terminal portion of the molecule in binding to MHC class II molecules and further indicate an important role for residues in other regions, particularly the disulfide loop.
Komisar
J L
JL
Department of Experimental Pathology, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100.
Small-Harris
S
S
Tseng
J
J
D000595:Amino Acid Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004768:Enterotoxins / Q000737:chemistry / Q000378:metabolism*; D006684:HLA-DR Antigens / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D013329:Structure-Activity Relationship; D018089:Superantigens / Q000737:chemistry / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004768:Enterotoxins; D006684:HLA-DR Antigens; D010455:Peptides; D018089:Superantigens; C031818:enterotoxin B, staphylococcal