Abstract
Gly m Bd 28 K is a major soybean (Glycine max Merr.) glycoprotein allergen. It was originally identified as a 28 kDa polypeptide in soybean seed flour. However, the full-length protein is encoded by an open reading frame (ORF) of 473 amino acids, and contains a 23 kDa C-terminal polypeptide of as yet unknown allergenic and structural characteristics. IgE-binding (allergenic potential) of the Gly m Bd 28 K protein including the 23 kDa C-terminal portion as well as shorter fragments derived from the full-length ORF were evaluated using sera from soy-sensitive adults. All of these sera contained IgE that efficiently recognized the C-terminal region. Epitope mapping demonstrated that a dominant linear C-terminal IgE binding epitope resides between residues S256 and A270. Alanine scanning of this dominant epitope indicated that five amino acids, Y260, D261, D262, K264 and D266, contribute most towards IgE-binding. A model based on the structure of the β subunit of soybean β-conglycinin revealed that Gly m Bd 28 K contains two cupin domains. The dominant epitope is on the edge of the first β-sheet of the C-terminal cupin domain and is present on a potentially solvent-accessible loop connecting the two cupin domains. Thus, the C-terminal 23 kDa polypeptide of Gly m Bd 28 K present in soy products is allergenic and apparently contains at least one immunodominant epitope near the edge of a cupin domain. This knowledge could be helpful in the future breeding of hypoallergenic soybeans.
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Abbreviations
- Ara h 1 :
-
Arachis hypogaea allergen 1
- Ara h 3 :
-
Arachis hypogaea allergen 3
- BCA :
-
Bicinchoninic acid
- Gly m Bd 28 K :
-
Glycine max band 28 kDa allergen
- Gly m Bd 30 K :
-
Glycine max band 30 kDa allergen
- Gly m Bd 68 K :
-
Glycine max band 68 kDa allergen
- IgE :
-
Immunoglobulin E
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Acknowledgements
We thank Dr. Rudolf Jung at Pioneer-Hybrid International for the gift of the soybean Gly m Bd 28 K clone. This work was supported in part by NIH Grant Number 1 P20 RR16469 from the BRIN Program of the National Center for Research Resources (G.S.). This work is published as Journal Series No. 14,087 from the Agriculture Research Division, University of Nebraska.
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Xiang, P., Haas, E.J., Zeece, M.G. et al. C-Terminal 23 kDa polypeptide of soybean Gly m Bd 28 K is a potential allergen. Planta 220, 56–63 (2004). https://doi.org/10.1007/s00425-004-1313-7
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DOI: https://doi.org/10.1007/s00425-004-1313-7