Antigenic specificity of a rabbit antiserum raised against the 15-28 segment of thymosin alpha 1.
Thymosin alpha 1, an acidic 28-residue peptide, enhances immune function. We have described a radioimmunoassay for this thymic factor based on a rabbit antiserum raised against a thymosin alpha 1-(15-28) conjugate (Incefy et al., J. Immun. Meth. 1986, in press). The detailed antigenic specificity of this antiserum was determined by measuring the ability of synthetic segments and analogues of thymosin alpha 1 and related peptides to compete with radioiodinated Ac-Tyr-thymosin alpha 1-(15-28) in this radioimmunoassay. The antiserum bound segments Ac-(1-28), (15-28), (20-28) and (21-28) with nearly equal efficiency but failed to bind segments Ac-(1-10), (11-20), (19-24) and (22-28). Thus, the major immunoreactive site seen by the antiserum is the COOH-terminal segment (21-28) (Glu-Val-Val-Glu-Glu-Ala-Glu-Asn-OH). Immunoreactivity of (21-28) was nearly abolished when the carboxylate groups of Glu-21, Glu-27 and Asn-28 were omitted separately. The antiserum bound to prothymosin alpha and thymosin alpha 11, which lack the alpha-carboxylate group of Asn-28, with 0.9 and 0.2%, respectively, of the efficiency of thymosin alpha 1. But it bound nonspecifically to parathymosin alpha, which contains the internal segment . . . -Glu-Val-Val-Glu-Glu-Glu-Glu-Asn- . . . . Residues Glu-21, Glu-27 and Asn-28 of thymosin alpha 1 may be important features of the antigenic site through their ability to induce helical structure, through the ability of their negatively charged carboxylate groups to bind to specific sites on the antibody or both.
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D000939:Epitopes / Q000032:analysis*; D007106:Immune Sera / Q000276:immunology; D010455:Peptides / Q000276:immunology; D011817:Rabbits; D011863:Radioimmunoassay; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000276:immunology
D000939:Epitopes; D007106:Immune Sera; D010455:Peptides; D013947:Thymosin; D000077596:Thymalfasin