Mapping of Malus domestica allergens by 2-D electrophoresis and IgE-reactivity

Anita Herndl, Gorji Marzban, Daniel Kolarich, Rainer Hahn, Donato Boscia, Wolfgang Hemmer, Fatemeh Maghuly, Elitza Stoyanova, Hermann Katinger, Margit Laimer*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    51 Citations (Scopus)

    Abstract

    The importance of apple allergens has been repeatedly emphasized, and their presence has been confirmed both in pollen and in fruits. In the present study, a combination of proteomic tools have been used to build a complete allergen map of apple. The water-soluble fraction of an apple extract was precipitated using a phenol-based procedure and separated by 2-DE. Initially four previously classified allergens, Mal d 1, Mal d 2, Mal d 3 and Mal d 4, could be identified in Western blots with polyclonal rabbit antibodies directed to the four respective allergens, and subsequently matched to the bands recognized by several patient sera. Further, all four known apple allergens were localized on a 2-DE map and they were matched with spots recognized by sera of patients with different allergic patterns. Moreover, a new, putative allergen could be identified using MS. We evaluated the influence of post-translational modifications and the immunoreactivity under different analytical conditions. The comparison of different visualization methods for 2-DE gels and blots revealed that even very low concentrations of the intact epitopes are detectable by IgEs of patients, and therefore might be sufficient to trigger allergic symptoms in sensitized individuals.

    Original languageEnglish
    Pages (from-to)437-448
    Number of pages12
    JournalElectrophoresis
    Volume28
    Issue number3
    DOIs
    Publication statusPublished - Feb 2007

    Keywords

    • Apple allergen map
    • Apple proteins
    • Fruit allergens
    • Mal d 1-4
    • Pathogenesis-related proteins

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