Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo
Background: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare...
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| Format: | Article |
| Language: | English |
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Varastegan Institute for Medical Sciences
2013-05-01
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| Series: | Reports of Biochemistry and Molecular Biology |
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| Online Access: | http://rbmb.net/article-1-30-en.pdf |
| _version_ | 1818540234481074176 |
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| author | Mojtaba Sankian Mahmoud Mahmoudi Abdol-Reza Varasteh |
| author_facet | Mojtaba Sankian Mahmoud Mahmoudi Abdol-Reza Varasteh |
| author_sort | Mojtaba Sankian |
| collection | DOAJ |
| description | Background: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare the IgE-binding capacity of the natural protein with the recombinant allergen, and to determine cross-reactivity of the major allergen with closely-related allergens from other plants displaying clinical cross-reactivity with melon.
Methods: Identification and molecular characterization of the major melon allergen were performed using IgE immunoblotting, allergen-specific ELISA, affinity-based purifications, cross-inhibition assays, cloning, and expression of the allergen in Escherichia coli.
Results: Melon profilin was identified and isolated as a major IgE-binding component and designated as Cuc m 2. Sequencing corresponding cDNA revealed an open reading frame of 363 bp coding for 131 amino acid residues and two fragments of 171 bp and 383 bps for the 5’and 3’ UTRs, respectively. Significant cross-reactivity was found between melon profilin and Cynodon dactylon, tomato, peach, and grape profilins in cross-inhibition assays. Although the highest degree of amino acid identity was revealed with watermelon profilin, there was no significant cross-reactivity between melon and watermelon profilins.
Conclusion: Melon profilin is the major IgE-binding component in melon extract, and the recombinant and natural forms exhibited similar IgE-binding capacities. A part of the fruit-fruit and pollen-fruit cross-reactions could be explained by the presence of this conserved protein; however, sequence homology provides insufficient information to predict IgE cross-reactivity of profilins. |
| first_indexed | 2024-12-11T21:52:38Z |
| format | Article |
| id | doaj.art-0192fd5836da4699975ac95b9ee2300b |
| institution | Directory Open Access Journal |
| issn | 2322-3480 2322-3480 |
| language | English |
| last_indexed | 2024-12-11T21:52:38Z |
| publishDate | 2013-05-01 |
| publisher | Varastegan Institute for Medical Sciences |
| record_format | Article |
| series | Reports of Biochemistry and Molecular Biology |
| spelling | doaj.art-0192fd5836da4699975ac95b9ee2300b2022-12-22T00:49:24ZengVarastegan Institute for Medical SciencesReports of Biochemistry and Molecular Biology2322-34802322-34802013-05-01124963Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis meloMojtaba Sankian0Mahmoud Mahmoudi1Abdol-Reza Varasteh2Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IranImmunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IranAllergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IranBackground: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare the IgE-binding capacity of the natural protein with the recombinant allergen, and to determine cross-reactivity of the major allergen with closely-related allergens from other plants displaying clinical cross-reactivity with melon. Methods: Identification and molecular characterization of the major melon allergen were performed using IgE immunoblotting, allergen-specific ELISA, affinity-based purifications, cross-inhibition assays, cloning, and expression of the allergen in Escherichia coli. Results: Melon profilin was identified and isolated as a major IgE-binding component and designated as Cuc m 2. Sequencing corresponding cDNA revealed an open reading frame of 363 bp coding for 131 amino acid residues and two fragments of 171 bp and 383 bps for the 5’and 3’ UTRs, respectively. Significant cross-reactivity was found between melon profilin and Cynodon dactylon, tomato, peach, and grape profilins in cross-inhibition assays. Although the highest degree of amino acid identity was revealed with watermelon profilin, there was no significant cross-reactivity between melon and watermelon profilins. Conclusion: Melon profilin is the major IgE-binding component in melon extract, and the recombinant and natural forms exhibited similar IgE-binding capacities. A part of the fruit-fruit and pollen-fruit cross-reactions could be explained by the presence of this conserved protein; however, sequence homology provides insufficient information to predict IgE cross-reactivity of profilins.http://rbmb.net/article-1-30-en.pdfCross-reactivityFruit allergyMelonProfilinRecombinant allergen |
| spellingShingle | Mojtaba Sankian Mahmoud Mahmoudi Abdol-Reza Varasteh Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo Reports of Biochemistry and Molecular Biology Cross-reactivity Fruit allergy Melon Profilin Recombinant allergen |
| title | Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo |
| title_full | Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo |
| title_fullStr | Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo |
| title_full_unstemmed | Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo |
| title_short | Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo |
| title_sort | molecular cloning characterization and expression of cuc m 2 a major allergen in cucumis melo |
| topic | Cross-reactivity Fruit allergy Melon Profilin Recombinant allergen |
| url | http://rbmb.net/article-1-30-en.pdf |
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