Molecular Characterization of Recombinant Mus a 5 Allergen from Banana Fruit |
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Authors: | Ivan Mrkic Mohamed Abughren Jasna Nikolic Uros Andjelkovic Emilia Vassilopoulou Athanassios Sinaniotis Arnd Petersen Nikolaos G. Papadopoulos Marija Gavrovic-Jankulovic |
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Affiliation: | 1. Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000, Belgrade, Serbia 2. Department of Chemistry, Institute for Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, Serbia 3. Allergy Research Centre, 2nd Department of Pediatrics, University of Athens, Athens, Greece 4. Allergy Department, “Sotiria” General Hospital, Athens, Greece 5. Division Clinical and Molecular Allergology, Research Center Borstel, Airway Research Center North (ARCN), Member of the German Center for Lung Research, Borstel, Germany
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Abstract: | Allergy to banana fruit appears to have become an important cause of fruit allergy in Europe. Among five allergens that have been found, beta-1,3-glucanase denoted as Mus a 5 was identified as a candidate allergen for the component-resolved allergy diagnosis of banana allergy. Because of the variations in protein levels in banana fruit, in this study Mus a 5 was produced as a fusion protein with glutathione-S-transferase in Escherichia coli. The recombinant Mus a 5 was purified under native conditions by a combination of affinity, ion-exchange, and reversed phase chromatography. N-terminal sequence was confirmed by Edman degradation and 55 % of the primary structure was identified by mass fingerprint, while the secondary structure was assessed by circular dichroism spectroscopy. IgG reactivity of recombinant protein was shown in 2-D immunoblot with anti-Mus a 5 antibodies, while IgG and IgE binding to natural Mus a 5 was inhibited with the recombinant Mus a 5 in immunoblot inhibition test. IgE reactivity of recombinant Mus a 5 was shown in ELISA within a group of ten persons sensitized to banana fruit. Recombinant Mus a 5 is a novel reagent suitable for the component-resolved allergy diagnosis of banana allergy. |
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