Molecular cloning and characterization of Cup a 4, a new allergen from Cupressus arizonica |
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Authors: | Yago Pico de Coaña Nuria Parody Miguel Ángel Fuertes Daniela Roncarolo Joaquín Sastre Carlos Alonso |
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Institution: | a Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain b R&D Department, Laboratorios LETI S.L., Madrid, Spain c Lofarma S.p.A., Milan, Italy d Allergy Unit, Bordighera Hospital, Imperia, Italy e Fundación Jiménez Díaz, Servicio de Alergia, Madrid, Spain |
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Abstract: | Sensitization to Cupressaceae pollen has become one of the most important causes of pollinosis in Western countries during winter and early spring. However, the characterization of the extracts, the allergens involved and the cross-reactivity with other pollen sources still remain poorly studied; in the case of Cupressus arizonica only two allergens have been described so far. A new allergen from C. arizonica pollen, Cup a 4, was cloned and expressed in Escherichia coli as an N-terminally His-tag recombinant protein that was characterized biochemically, immunologically and by circular dichroism spectroscopy. The new allergen has high sequence identity with Prickly Juniper allergen Jun o 4 and contains four EF-hand domains. The recombinant protein has structural similarities with other calcium binding allergens such as Ole e 3, Ole e 8 and Phl p 7. Cup a 4 is expressed in mature pollen grains and shares antigenic properties with the recombinant form. Sera from 9.6% C. arizonica allergic patients contain specific IgE antibodies against recombinant Cup a 4. |
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Keywords: | Cypress pollen Allergen Polcalcin EF-hand Recombinant |
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