Structure-activity relationship of toxic-shock-syndrome toxin-1: derivation and characterization of immunologically and biologically active fragments.
Toxic-shock-syndrome toxin-1 (TSST-1), a 22-kilodalton (kDa) polypeptide, was proteolyzed by papain, generating three distinct fragments, identified as 16, 12, and 10 kDa (based on molecular masses estimated from the predicted amino acid sequence). The NH2-terminal sequence analysis of the fragments indicated that the peptide bonds between Tyr-52 and Ser-53 and between Gly-87 and Val-88 were cleaved. Functional activity, evaluated through enzyme-linked immunosorbent and inhibition assays, was demonstrated only with the 16- and 12-kDa fragments. The presence of homologous and heterologous antigenic determinants on the fragments was demonstrated by immunoblotting. In in vitro stimulation of human peripheral blood mononuclear cells, the 12-kDa fragment was significantly (P = .003) more active than the 16-kDa fragment. The former composed 75% of the latter and occupied the COOH-terminal portion of the holotoxin. The functional domains were located on two-thirds of the TSST-1 molecule, toward the COOH-terminal end, and mitogenicity apparently was separable from serological activity.
Edwin
C
C
Channing Laboratory, Boston, Massachusetts 02115.
Parsonnet
J
J
Kass
E H
EH
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001427:Bacterial Toxins; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010206:Papain / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D013211:Staphylococcus aureus; D013329:Structure-Activity Relationship; D018089:Superantigens
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D004768:Enterotoxins; D010446:Peptide Fragments; D018089:Superantigens; C030107:enterotoxin F, Staphylococcal; D010206:Papain