N-terminal amino acids of bovine alpha interferons are relevant for the neutralization of their antiviral activity.
The structure-function relationship of interferons (IFNs) has been studied by epitope mapping. Epitopes of bovine IFNs, however, are practically unknown, despite their importance in virus infections and in the maternal recognition of pregnancy. It has been shown that recombinant bovine (rBo)IFN-alphaC and rBoIFN-alpha1 differ only in 12 amino acids and that the F12 monoclonal antibody (mAb) binds to a linear sequence of residues 10 to 34. We show here that the antiviral activities of these two IFNs were neutralized by the F12 mAb to different extents using two tests. In residual activity tests the antiviral activity dropped by more than 99% with rBoIFN-alphaC and by 84% with rBoIFN-alpha1. In checkerboard antibody titrations, the F12 mAb titer was 12,000 with rBoIFN-alphaC and only 600 with rBoIFN-alpha1. Since these IFNs differ in their amino acid sequence at positions 11, 16 and 19 of the amino terminus, only these amino acids could account for the different neutralization titers, and they should participate in antibody binding. According to the three-dimensional structure described for human and murine IFNs, these amino acids are located in the alpha helix A; amino acids 16 and 19 of the bovine IFNs would be expected to be exposed and could bind to the antibody directly. The amino acid at position 11 forms a hydrogen bond in human IFNs-alpha and it is possible that, in bovine IFNs-alpha, the F12 mAb, binding near position 11, would disturb this hydrogen bond, resulting in the difference in the extent of neutralization observed.
Barreto Filho
J B
JB
Departamento de Medicina Veterinária, Universidade Federal de Lavras, Lavras, MG, Brasil.
Eiras
P R
PR
Golgher
R R
RR
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000998:Antiviral Agents / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics; D002417:Cattle; D000939:Epitopes; D006801:Humans; D007370:Interferon Type I / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics; D009500:Neutralization Tests; D011994:Recombinant Proteins
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D000939:Epitopes; D007370:Interferon Type I; D011994:Recombinant Proteins