Molecular and structural analysis of a continuous birch profilin epitope defined by a monoclonal antibody

被引:35
作者
Wiedemann, P
Giehl, K
Almo, SC
Fedorov, AA
Girvin, M
Steinberger, P
Rudiger, M
Ortner, M
Sippl, M
Dolecek, C
Kraft, D
Jockusch, B
Valenta, R
机构
[1] UNIV VIENNA,AKH,INST GEN & EXPT PATHOL,A-1090 VIENNA,AUSTRIA
[2] TECH UNIV CAROLO WILHELMINA BRAUNSCHWEIG,INST ZOOL,DEPT CELL BIOL,D-38106 BRAUNSCHWEIG,GERMANY
[3] ALBERT EINSTEIN COLL MED,DEPT BIOCHEM,BRONX,NY 10461
[4] SALZBURG UNIV,CTR APPL MOL ENGN,A-5020 SALZBURG,AUSTRIA
关键词
D O I
10.1074/jbc.271.47.29915
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 amino acid (Gln-47 --> Glu) within the epitope was found to abolish the bind ing of monoclonal antibody 4A6 to other plant profilins. The MMR 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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页码:29915 / 29921
页数:7
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