Epitope mapping of the major allergen from yellow mustard seeds, Sin a I.
The antigenic sites on the major allergen from yellow mustard (Sinapis alba L.) seeds were studied using murine (BALB/c) monoclonal antibodies (mAb) and human IgE antibodies. Ten IgG1 (K) mAb from two fusions were analyzed. Competition and complementation studies performed with peroxidase labeled mAb reveal the existence of two main antigenic sites in Sin a I. All the described mAb failed to recognize the unordered carboxyamidomethylated polypeptide chains, with the single exception of 2B3, which binds the alkylated large chain. However, this mAB cannot react with the tetranitromethane-modified protein which retains the native conformation. This fact suggests that the only tyrosine of Sin a I, located in the large chain, may be part of a sequential epitope of the allergen. This chemical modification also alters the binding of the mAb 4A11 and 3F3 to the allergen, besides 2B3. The three mAb belong to the same complementation group. Specific IgE binding cannot be inhibited either by the large or small carboxyamidomethylated polypeptide chains, while the nitrated allergen shows a weaker inhibitory activity than the native Sin a I. 4A11, which is a tyrosine-dependent mAb, causes the greatest binding inhibition of the tested mAb on human IgE from atopic individuals, as determined from a reverse enzyme immunoassay, suggesting an important role played by tyrosine in the immunochemical recognition of Sin a I.
Departamento de Bioquímica y Biología Molecular I, Facultad de Ciencias, Universidad Complutense, Madrid, Spain.
L
L
Menéndez-Arias
J
J
Domínguez
I
I
Moneo
R
R
Rodríguez
D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001937:Brassica / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D009149:Mustard Plant / Q000276:immunology*; D010946:Plants, Medicinal; D012639:Seeds / Q000276:immunology*
D000485:Allergens; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007073:Immunoglobulin E