Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family

Fang Wang, Jason M. Robotham, Suzanne S Teuber, Shridhar K. Sathe, Kenneth H. Roux

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Background: We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. Methods: A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. Results: Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew-allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. Conclusions: We conclude that this legumin-like protein is a major allergen in cashew nut.

Original languageEnglish (US)
Pages (from-to)27-39
Number of pages13
JournalInternational Archives of Allergy and Immunology
Volume132
Issue number1
DOIs
StatePublished - 2003

Fingerprint

Anacardium
Nuts
Allergens
Peptides
Immunoblotting
Immunoglobulin E
Clone Cells
Epitopes
Serum
Seed Storage Proteins
Epitope Mapping
Escherichia coli Proteins
Globulins
Gene Library
Soybeans
Sequence Analysis
Immune Sera
Albumins
Rabbits
Technology

Keywords

  • 11S globulin
  • Ana o 2
  • Cashew allergen
  • Food allergy
  • Legumin-like protein
  • Seed storage proteins
  • Tree nut allergy

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family. / Wang, Fang; Robotham, Jason M.; Teuber, Suzanne S; Sathe, Shridhar K.; Roux, Kenneth H.

In: International Archives of Allergy and Immunology, Vol. 132, No. 1, 2003, p. 27-39.

Research output: Contribution to journalArticle

Wang, Fang ; Robotham, Jason M. ; Teuber, Suzanne S ; Sathe, Shridhar K. ; Roux, Kenneth H. / Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family. In: International Archives of Allergy and Immunology. 2003 ; Vol. 132, No. 1. pp. 27-39.
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AU - Roux, Kenneth H.

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N2 - Background: We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. Methods: A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. Results: Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew-allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. Conclusions: We conclude that this legumin-like protein is a major allergen in cashew nut.

AB - Background: We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. Methods: A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. Results: Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew-allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. Conclusions: We conclude that this legumin-like protein is a major allergen in cashew nut.

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KW - Seed storage proteins

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