An experimental and modeling-based approach to locate IgE epitopes of plant profilin allergens Gema López-Torrejón, PhD, Araceli Díaz-Perales, PhD, Julia.

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Presentation transcript:

An experimental and modeling-based approach to locate IgE epitopes of plant profilin allergens Gema López-Torrejón, PhD, Araceli Díaz-Perales, PhD, Julia Rodríguez, MD, PhD, Rosa Sánchez-Monge, PhD, Jesus F. Crespo, MD, PhD, Gabriel Salcedo, PhD, Luis F. Pacios, PhD Journal of Allergy and Clinical Immunology Volume 119, Issue 6, Pages 1481-1488 (June 2007) DOI: 10.1016/j.jaci.2007.02.004 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, Ribbon diagram of Cuc m 2. B, Profile of percentage of solvent exposure. C, Surface at the orientation in Fig 1, A (left), and after a counterclockwise 120° rotation about a vertical axis (right) showing (1) PLP-binding (green) and (2) actin-binding (red) sites and (3) plant-specific pocket (blue). Overlap between 2 and 3 is colored purple. D, PB potential mapped onto surfaces. Journal of Allergy and Clinical Immunology 2007 119, 1481-1488DOI: (10.1016/j.jaci.2007.02.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 A, Protein staining and IgE immunodetection with sera from patients with melon allergy of purified natural (n) and recombinant (r) Cuc m 2 and ovalbumin (negative control; C). B, Specific IgE of 10 individual sera. BSA (C) was tested as a negative control. Means (n = 3) and SDs (bars) are represented. OD values of greater than 0.24 units (3 × 0.081 of the mean [n = 7] obtained for the negative control) were considered positive results. Journal of Allergy and Clinical Immunology 2007 119, 1481-1488DOI: (10.1016/j.jaci.2007.02.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 A, IgE immunodetection of SPOTs with synthetic peptides spanning the entire Cuc m 2 amino acid sequence by using a serum pool and 4 individual sera (numbers as in Fig 2, B) from patients with melon allergy. B, Amino acid sequence highlighting the positions of major (bold and shaded) and minor (bold) reactive peptides to the serum pool or the individual sera. Journal of Allergy and Clinical Immunology 2007 119, 1481-1488DOI: (10.1016/j.jaci.2007.02.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 A-C, Epitopes corresponding to peptides 14 (green), 17 to 19 (blue), and 26 (magenta). Fig 4, A, Ribbon diagrams; Fig 4, B, surfaces; and Fig 4, C, overlap with actin-binding site (red). Bottom views of Fig 4, A through C, are derived from upper views by means of a clockwise 90° rotation about a vertical axis (arrows point to the plant-specific pocket). D and E, Ribbon diagram (Fig 4, D) and surface (Fig 4, E) of theoretical epitopes E3 (yellow) and E4 (cyan). Journal of Allergy and Clinical Immunology 2007 119, 1481-1488DOI: (10.1016/j.jaci.2007.02.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 A, Surfaces of epitopes E1 (blue) and E2 (magenta). B, Isocontours −2.0 (red) and +2.0 (blue) of PB potential. C, PB potential mapped onto the surface of E1 and E2. D, E1 in front. E, E2 in front. F, Surfaces of E3 (yellow) and E4 (cyan) after a 180° rotation of Fig 5, A, about a vertical axis. G, PB potential. Journal of Allergy and Clinical Immunology 2007 119, 1481-1488DOI: (10.1016/j.jaci.2007.02.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions