Molecular basis of allergen cross-reactivity: non-specific lipid transfer proteins from wheat flour and peach fruit as models.
Peach non-specific lipid transfer protein (Pru p 3; nsLTP) has been characterized as the major food allergen in the adult Mediterranean population. Its wheat homologous protein, Tri a 14 has a relevant inhalant allergen in occupational baker's asthma. Different sensitization patterns to these allergens have been found in patients with this latter disorder. The objective of the present study was to characterize IgE epitopes of Tri a 14 and to compare them with those of Pru p 3 using three complementary strategies: the analysis of IgE-binding capacity of decapeptides bound to membrane, the identification of mimotopes using a phage display random peptide library, and the analysis of the surface electrostatic potential of both allergens. Thus, synthetic overlapping decapeptides, covering the Pru p 3 and Tri a 14 amino acid sequences, were used to identify sequential regions involved in recognition of IgE from baker's asthma patients sensitized to both nsLTPs. A phage display library was screened with total IgE from the same patients, and positive clones sequentially selected using the purified allergens, allowed to identify mimotopes (conformational epitopes) of Tri a 14 and Pru p 3. Both sequential regions and mimotopes were localized in the corresponding 3D molecular surface and their electrostatic properties were analyzed. Common sequential regions with strong IgE-binding capacity (residues 31-40 and 71-80) were identified in Tri a 14 and Pru p 3, whereas regions Tri a 14(51-60) and Pru p 3(11-20) were found specific of each allergen. A major conformational epitope (mimotope), L34H35N36R39S40S42D43G74V75L77P78Y79T80, which comprised the two common sequential epitopes, was located in Tri a 14, and a very similar one in Pru p 3. However, differences were detected on the surface electrostatic potential of both mimotopes: a first part (around residues 31-45) showed similar positive features in both allergens, whereas a second part (around residues 74-80) was markedly negative in Tri a 14 but neutral-positive in Pru p 3. Tri a 14 and Pru p 3 have a similar conformational region involved in IgE-binding, although their electrostatic features are different. Additionally, common and specific sequential IgE-binding regions were mapped in both allergens. These findings could be instrumental in understanding the cross-reactivity and specificity of sensitization to both homologous allergens.
Unidad de Bioquímica, Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, UPM, Ciudad Universitaria, 28040 Madrid, Spain.
Leticia
L
Tordesillas
Luis F
LF
Pacios
Arancha
A
Palacin
Santiago
S
Quirce
Alicia
A
Armentia
Domingo
D
Barber
Gabriel
G
Salcedo
Araceli
A
Diaz-Perales
D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology*; D002999:Clone Cells; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology; D005433:Flour; D005638:Fruit / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D018448:Models, Immunological; D008958:Models, Molecular; D018716:Molecular Mimicry / Q000276:immunology; D008969:Molecular Sequence Data; D027861:Prunus / Q000276:immunology*; D014908:Triticum / Q000276:immunology*
D000485:Allergens; D052179:Antigens, Plant; D002352:Carrier Proteins; D000939:Epitopes; C072087:sterol carrier proteins; D007073:Immunoglobulin E