TY - JOUR
T1 - Molecular and structural analysis of a continuous birch profilin epitope defined by a monoclonal antibody
AU - Wiedemann, Petra
AU - Giehl, Klaudia
AU - Almo, Steven C.
AU - Fedorov, Alexander A.
AU - Girvin, Mark
AU - Steinberger, Peter
AU - Rüdiger, Manfred
AU - Ortner, Maria
AU - Sippl, Manfred
AU - Dolecek, Christiane
AU - Kraft, Dietrich
AU - Jockusch, Brigitte
AU - Valenta, Rudolf
PY - 1996
Y1 - 1996
N2 - The interaction of a mouse monoclonal antibody (4A6) and birch profilin, a structurally well conserved actin- and phosphoinositide-binding protein and cross-reactive allergen, was characterized. In contrast to serum IgE from allergic patients, which shows cross-reactivity with most plants, monoclonal antibody 4A6 selectively reacted with tree pollen profilins. Using synthetic overlapping peptides, a continuous hexapeptide epitope was identified. The exchange of a single amine acid (Gln-47→Glu) within the epitope was found to abolish the binding of monoclonal antibody 4A6 to other plant profilins. The NMR analyses of the birch and the nonreactive timothy grass profilin peptides showed that the loss of binding was not due to major structural differences. Both peptides adopted extended conformations similar to that observed for the epitope in the x-ray crystal structure of the native birch profilin. Binding studies with peptides and birch profilin mutants generated by in vitro mutagenesis demonstrated that the change of Gln-47 to acidic amino acids (e.g. Glu or Asp) led to electrostatic repulsion of monoclonal antibody 4A6. In conclusion the molecular and structural analyses of the interaction of a monoclonal antibody with a continuous peptide epitope, recognized in a conformation similar to that displayed on the native protein, are presented.
AB - The interaction of a mouse monoclonal antibody (4A6) and birch profilin, a structurally well conserved actin- and phosphoinositide-binding protein and cross-reactive allergen, was characterized. In contrast to serum IgE from allergic patients, which shows cross-reactivity with most plants, monoclonal antibody 4A6 selectively reacted with tree pollen profilins. Using synthetic overlapping peptides, a continuous hexapeptide epitope was identified. The exchange of a single amine acid (Gln-47→Glu) within the epitope was found to abolish the binding of monoclonal antibody 4A6 to other plant profilins. The NMR analyses of the birch and the nonreactive timothy grass profilin peptides showed that the loss of binding was not due to major structural differences. Both peptides adopted extended conformations similar to that observed for the epitope in the x-ray crystal structure of the native birch profilin. Binding studies with peptides and birch profilin mutants generated by in vitro mutagenesis demonstrated that the change of Gln-47 to acidic amino acids (e.g. Glu or Asp) led to electrostatic repulsion of monoclonal antibody 4A6. In conclusion the molecular and structural analyses of the interaction of a monoclonal antibody with a continuous peptide epitope, recognized in a conformation similar to that displayed on the native protein, are presented.
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U2 - 10.1074/jbc.271.47.29915
DO - 10.1074/jbc.271.47.29915
M3 - Article
C2 - 8939935
AN - SCOPUS:10544242206
SN - 0021-9258
VL - 271
SP - 29915
EP - 29921
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 47
ER -