Identification of IgE-binding epitopes of the major peach allergen Pru p 3

García‐Casado, Gloria; Pacios, Luis F.; Díaz‐Perales, Araceli; Sánchez‐Monge, Rosa; Lombardero, M.; García-Sellés, F J; Polo, F.; Barber, Domingo; Salcedo, Gabriel

doi:10.1016/s0091-6749(03)01605-1

Cited by 120 publications

(103 citation statements)

References 23 publications

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“…The experimentally determined N-terminal amino acid sequence for rCit s 3.0102 is underlined. Sequential IgE epitopes of Pru p 3 previously described [24] are in boxes. The additional N-terminus (E) of the recombinant allergens was added for their proper expression in P. pastoris .…”

Section: Resultsmentioning

confidence: 99%

Lipid Transfer Proteins and Allergy to Oranges

Ahrazem¹,

Ibáñez

López‐Torrejón³

et al. 2005

Int Arch Allergy Immunol

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Background: Lipid transfer proteins (LTPs) are relevant fruit allergens, recently proposed as model plant food allergens. No citrus fruit allergen has been characterized to date. We sought to identify and isolate citrus fruit LTPs and to explore their relevance in orange allergy. Methods: Twenty-seven patients, showing mainly oral allergy syndrome after orange ingestion, as well as positive prick responses and serum-specific IgE levels to orange, were selected. Natural orange and lemon LTPs, as well as a recombinant orange LTP isoform expressed in Pichia pastoris, were isolated by chromatographic methods and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionizaion mass spectrometry, and DNA sequencing of the corresponding cDNA in the case of the recombinant allergen. Specific IgE determination, immunodetection, ELISA-inhibition assays and in vivo skin prick tests (SPTs) were performed with all three purified allergens and with the major peach LTP allergen, Pru p 3. Results: The natural allergens purified from orange (nCit s 3) and lemon (nCit l 3) showed very similar N-terminal amino acid sequences (18 out of 20 identical residues), typical of LTPs, and molecular masses of 9,610 and 9,618 Da, respectively. The recombinant orange isoform (rCit s 3) expressed in P. pastoris (16 out of 20 residues identical to its natural counterpart in the N-terminal region) presented 92 amino acid residues and 9,463 Da, and 67% sequence identity with rPru p 3. Of the 27 sera analyzed, specific IgE to the purified allergens was found in 54% for nCit l 3, 48% for nCit s 3, 46% for rCit s 3 and 37% for rPru p 3. Positive SPT responses were obtained in 7 out of 26 patients tested for nCit s 3, 3 out of 8 for nCit l 3 and 10 out of 26 for nPru p 3. ELISA-inhibition assays showed an equivalent IgE-binding pattern for the natural and recombinant orange LTPs, and IgE cross-reactivity among the purified orange, lemon and peach LTP allergens. Conclusions: Members of the LTP allergen family are involved in allergy to oranges, displaying positive in vitro and in vivo reactions in 30–50% of the patients studied. Both orange and lemon allergens show cross-reactivity with the major peach allergen Pru p 3.

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Section: Resultsmentioning

confidence: 99%

Lipid Transfer Proteins and Allergy to Oranges

Ahrazem¹,

Ibáñez

López‐Torrejón³

et al. 2005

Int Arch Allergy Immunol

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“…Recently reported evidence on the possible multifunctionality of nsLTPs in plant growth processes [6][7][8] and open questions on the antigenic interactions of plant nsLTPs in alimentary allergies (particularly peach nsLTP), [58][59][60][61] pose the need to deepen our understanding on their lipid binding properties. In addition, we aimed to gain insight into general protein-ligand interactions by providing quantitative information on the lipid binding abilities of these small proteins.…”

Section: Discussionmentioning

confidence: 99%

Computational study of ligand binding in lipid transfer proteins: Structures, interfaces, and free energies of protein‐lipid complexes

et al. 2012

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Plant nonspecific lipid transfer proteins (nsLTPs) bind a wide variety of lipids, which allows them to perform disparate functions. Recent reports on their multifunctionality in plant growth processes have posed new questions on the versatile binding abilities of these proteins. The lack of binding specificity has been customarily explained in qualitative terms on the basis of a supposed structural flexibility and nonspecificity of hydrophobic protein-ligand interactions. We present here a computational study of protein-ligand complexes formed between five nsLTPs and seven lipids bound in two different ways in every receptor protein. After optimizing geometries in molecular dynamics calculations, we computed PoissonBoltzmann electrostatic potentials, solvation energies, properties of the protein-ligand interfaces, and estimates of binding free energies of the resulting complexes. Our results provide the first quantitative information on the ligand abilities of nsLTPs, shed new light into protein-lipid interactions, and reveal new features which supplement commonly held assumptions on their lack of binding specificity.

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“…Garcia-Casado et al (2003) used the peach allergen, Pru p 3, as a model to find IgE epitopes. They identified three amino acid sequence regions (11-25, 31-45 and 71-80) as major IgE epitopes and predicted five amino acid residues of R39, T40, R44, K80 and K91 as antibody recognition sites using IgE immuno-detection and synthetic peptides.…”

Section: Discussionmentioning

confidence: 99%

Linkage map positions and allelic diversity of two Mal d 3 (non-specific lipid transfer protein) genes in the cultivated apple (Malus domestica)

et al. 2005

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Non-specific lipid transfer proteins (nsLTPs) of Rosaceae fruits, such as peach, apricot, cherry, plum and apple, represent major allergens for Mediterranean atopic populations. As a first step in elucidating the genetics of nsLTPs, we directed the research reported here towards identifying the number and location of nsLTP (Mal d 3) genes in the apple genome and determining their allelic diversity. PCR cloning was initially performed on two cultivars, Prima and Fiesta, parents of a core apple mapping progeny in Europe, based on two Mal d 3 sequences (AF221502 and AJ277164) in the GenBank. This resulted in the identification of two distinct sequences (representing two genes) encoding the mature nsLTP proteins. One is identical to accession AF221502 and has been named Mal d 3.01, and the other is new and has been named Mal d 3.02. Subsequent genome walking in the upstream direction and DNA polymorphism analysis revealed that these two genes are intronless and that they could be mapped on two homoeologous segments of linkage groups 12 and 4, respectively. Further cloning and sequencing of the coding and upstream region of both Mal d 3 genes in eight cultivars was performed to identify allelic variation. Assessment of the deduced nsLTP amino acid sequences gave a total of two variants at the protein level for Mal d 3.01 and three for Mal d 3.02. The consequences of our results for allergen nomenclature and the breeding of low allergenic apple cultivars are discussed.

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Identification of IgE-binding epitopes of the major peach allergen Pru p 3

Cited by 120 publications

References 23 publications

Lipid Transfer Proteins and Allergy to Oranges

Lipid Transfer Proteins and Allergy to Oranges

Computational study of ligand binding in lipid transfer proteins: Structures, interfaces, and free energies of protein‐lipid complexes

Linkage map positions and allelic diversity of two Mal d 3 (non-specific lipid transfer protein) genes in the cultivated apple (Malus domestica)

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