Mimotope mapping as a complementary strategy to define allergen IgE-epitopes: peach Pru p 3 allergen as a model.
Lipid transfer proteins (LTPs) are the major allergens of Rosaceae fruits in the Mediterranean area. Pru p 3, the LTP and major allergen of peach, is a suitable model for studying food allergy and amino acid sequences related with its IgE-binding capacity. In this work, we sought to map IgE mimotopes on the structure of Pru p 3, using the combination of a random peptide phage display library and a three-dimensional modelling approach. Pru p 3-specific IgE was purified from 2 different pools of sera from peach allergic patients grouped by symptoms (OAS-pool or SYS-pool), and used for screening of a random dodecapeptide phage display library. Positive clones were further confirmed by ELISA assays testing individual sera from each pool. Three-dimensional modelling allowed location of mimotopes based on analysis of electrostatic properties and solvent exposure of the Pru p 3 surface. Twenty-one phage clones were selected using Pru p 3-specific IgE, 9 of which were chosen using OAS-specific IgE while the other 12 were selected with systemic-specific IgE. Peptide alignments revealed consensus sequences for each pool: L37 R39 T40 P42 D43 R44 A46 P70 S76 P78 Y79 for OAS-IgE, and N35 N36 L37 R39 T40 D43 A46 S76 I77 P78 for systemic-IgE. These 2 consensus sequences were mapped on the same surface of Pru p 3, corresponding to the helix 2-loop-helix 3 region and part of the non-structured C-terminal coil. Thus, 2 relevant conformational IgE-binding regions of Pru p 3 were identified using a random peptide phage display library. Mimotopes can be used to study the interaction between allergens and IgE, and to accelerate the process to design new vaccines and new immunotherapy strategies.
Pacios
Luis F
LF
Unidad de Química y Bioquímica, Departamento de Biotecnología, E.T.S. Ingenieros de Montes, Madrid, Spain.
Tordesillas
Leticia
L
Cuesta-Herranz
Javier
J
Compes
Esther
E
Sánchez-Monge
Rosa
R
Palacín
Arantxa
A
Salcedo
Gabriel
G
Díaz-Perales
Araceli
A
D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D000918:Antibody Specificity / Q000276:immunology; D052179:Antigens, Plant; D001435:Bacteriophages; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D002999:Clone Cells; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D018448:Models, Immunological; D008958:Models, Molecular; D018716:Molecular Mimicry / Q000276:immunology*; D008969:Molecular Sequence Data; D010940:Plant Proteins; D017433:Protein Structure, Secondary; D027861:Prunus / Q000276:immunology*; D016415:Sequence Alignment
D000485:Allergens; D052179:Antigens, Plant; D002352:Carrier Proteins; D000939:Epitopes; D010940:Plant Proteins; C463181:Pru p 3 allergen; C030363:lipid transfer protein; D007073:Immunoglobulin E