Ana o 3, an important cashew nut (Anacardium occidentale L.) allergen of the 2S albumin family

Jason M. Robotham, Fang Wang, Vanessa Seamon, Suzanne S Teuber, Shridhar K. Sathe, Hugh A. Sampson, Kirsten Beyer, Margaret Seavy, Kenneth H. Roux

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

Background: Cashew nut allergy is the second most commonly reported tree nut allergy in the United States. We have previously cloned and characterized major cashew allergens belonging to the vicilin and legumin families of seed storage proteins. Objective: Here we set out to describe a third major cashew allergen, a 2S albumin. Methods: The recombinant cashew 2S albumin was amplified from a cDNA library by means of PCR, sequenced, and expressed in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition immunoblotting was used to identify the corresponding native cashew nut proteins. The mass of affinity-purified native allergen was determined by means of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy. Patients' sera were used to probe solid-phase 2S albumin peptides to identify linear epitopes. Results: The cloned allergen, designated Ana o 3, was identified as 2S albumin. MALDI-TOF mass spectroscopy of native Ana o 3 yielded a molecular mass of 12,598 d. Immunoblot analysis showed 21 (81%) of 26 sera from patients with cashew allergy were reactive. Three native Ana o 3 large-subunit isoforms with molecular weights ranging from approximately 6 to 10 kd were identified. Probing of overlapping synthetic Ana o 3 peptides with patients' sera identified 16 reactive peptides, 4 of which gave strong signals and one of which positionally overlaps linear epitopes in mustard and walnut allergenic 2S albumins. The overlapping cashew and walnut epitopes also share considerable homology. Conclusions: We conclude that this 2S albumin protein is a major allergen in cashew nut and demonstrates a possible basis for cross-reactivity with walnut 2S albumin.

Original languageEnglish (US)
Pages (from-to)1284-1290
Number of pages7
JournalJournal of Allergy and Clinical Immunology
Volume115
Issue number6
DOIs
StatePublished - Jun 2005

Fingerprint

Anacardium
Nuts
Allergens
Albumins
Juglans
Nut Hypersensitivity
Epitopes
Serum
Immunoblotting
Peptides
Mass Spectrometry
Lasers
Seed Storage Proteins
Mustard Plant
Gene Library
Hypersensitivity
Protein Isoforms
Proteins
Molecular Weight
Escherichia coli

Keywords

  • 2S albumin
  • Ana o 3
  • Cashew allergen
  • Epitope mapping
  • Food allergy
  • Seed storage proteins
  • Tree nut allergy

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Ana o 3, an important cashew nut (Anacardium occidentale L.) allergen of the 2S albumin family. / Robotham, Jason M.; Wang, Fang; Seamon, Vanessa; Teuber, Suzanne S; Sathe, Shridhar K.; Sampson, Hugh A.; Beyer, Kirsten; Seavy, Margaret; Roux, Kenneth H.

In: Journal of Allergy and Clinical Immunology, Vol. 115, No. 6, 06.2005, p. 1284-1290.

Research output: Contribution to journalArticle

Robotham, JM, Wang, F, Seamon, V, Teuber, SS, Sathe, SK, Sampson, HA, Beyer, K, Seavy, M & Roux, KH 2005, 'Ana o 3, an important cashew nut (Anacardium occidentale L.) allergen of the 2S albumin family', Journal of Allergy and Clinical Immunology, vol. 115, no. 6, pp. 1284-1290. https://doi.org/10.1016/j.jaci.2005.02.028
Robotham, Jason M. ; Wang, Fang ; Seamon, Vanessa ; Teuber, Suzanne S ; Sathe, Shridhar K. ; Sampson, Hugh A. ; Beyer, Kirsten ; Seavy, Margaret ; Roux, Kenneth H. / Ana o 3, an important cashew nut (Anacardium occidentale L.) allergen of the 2S albumin family. In: Journal of Allergy and Clinical Immunology. 2005 ; Vol. 115, No. 6. pp. 1284-1290.
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abstract = "Background: Cashew nut allergy is the second most commonly reported tree nut allergy in the United States. We have previously cloned and characterized major cashew allergens belonging to the vicilin and legumin families of seed storage proteins. Objective: Here we set out to describe a third major cashew allergen, a 2S albumin. Methods: The recombinant cashew 2S albumin was amplified from a cDNA library by means of PCR, sequenced, and expressed in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition immunoblotting was used to identify the corresponding native cashew nut proteins. The mass of affinity-purified native allergen was determined by means of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy. Patients' sera were used to probe solid-phase 2S albumin peptides to identify linear epitopes. Results: The cloned allergen, designated Ana o 3, was identified as 2S albumin. MALDI-TOF mass spectroscopy of native Ana o 3 yielded a molecular mass of 12,598 d. Immunoblot analysis showed 21 (81{\%}) of 26 sera from patients with cashew allergy were reactive. Three native Ana o 3 large-subunit isoforms with molecular weights ranging from approximately 6 to 10 kd were identified. Probing of overlapping synthetic Ana o 3 peptides with patients' sera identified 16 reactive peptides, 4 of which gave strong signals and one of which positionally overlaps linear epitopes in mustard and walnut allergenic 2S albumins. The overlapping cashew and walnut epitopes also share considerable homology. Conclusions: We conclude that this 2S albumin protein is a major allergen in cashew nut and demonstrates a possible basis for cross-reactivity with walnut 2S albumin.",
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T1 - Ana o 3, an important cashew nut (Anacardium occidentale L.) allergen of the 2S albumin family

AU - Robotham, Jason M.

AU - Wang, Fang

AU - Seamon, Vanessa

AU - Teuber, Suzanne S

AU - Sathe, Shridhar K.

AU - Sampson, Hugh A.

AU - Beyer, Kirsten

AU - Seavy, Margaret

AU - Roux, Kenneth H.

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N2 - Background: Cashew nut allergy is the second most commonly reported tree nut allergy in the United States. We have previously cloned and characterized major cashew allergens belonging to the vicilin and legumin families of seed storage proteins. Objective: Here we set out to describe a third major cashew allergen, a 2S albumin. Methods: The recombinant cashew 2S albumin was amplified from a cDNA library by means of PCR, sequenced, and expressed in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition immunoblotting was used to identify the corresponding native cashew nut proteins. The mass of affinity-purified native allergen was determined by means of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy. Patients' sera were used to probe solid-phase 2S albumin peptides to identify linear epitopes. Results: The cloned allergen, designated Ana o 3, was identified as 2S albumin. MALDI-TOF mass spectroscopy of native Ana o 3 yielded a molecular mass of 12,598 d. Immunoblot analysis showed 21 (81%) of 26 sera from patients with cashew allergy were reactive. Three native Ana o 3 large-subunit isoforms with molecular weights ranging from approximately 6 to 10 kd were identified. Probing of overlapping synthetic Ana o 3 peptides with patients' sera identified 16 reactive peptides, 4 of which gave strong signals and one of which positionally overlaps linear epitopes in mustard and walnut allergenic 2S albumins. The overlapping cashew and walnut epitopes also share considerable homology. Conclusions: We conclude that this 2S albumin protein is a major allergen in cashew nut and demonstrates a possible basis for cross-reactivity with walnut 2S albumin.

AB - Background: Cashew nut allergy is the second most commonly reported tree nut allergy in the United States. We have previously cloned and characterized major cashew allergens belonging to the vicilin and legumin families of seed storage proteins. Objective: Here we set out to describe a third major cashew allergen, a 2S albumin. Methods: The recombinant cashew 2S albumin was amplified from a cDNA library by means of PCR, sequenced, and expressed in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition immunoblotting was used to identify the corresponding native cashew nut proteins. The mass of affinity-purified native allergen was determined by means of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy. Patients' sera were used to probe solid-phase 2S albumin peptides to identify linear epitopes. Results: The cloned allergen, designated Ana o 3, was identified as 2S albumin. MALDI-TOF mass spectroscopy of native Ana o 3 yielded a molecular mass of 12,598 d. Immunoblot analysis showed 21 (81%) of 26 sera from patients with cashew allergy were reactive. Three native Ana o 3 large-subunit isoforms with molecular weights ranging from approximately 6 to 10 kd were identified. Probing of overlapping synthetic Ana o 3 peptides with patients' sera identified 16 reactive peptides, 4 of which gave strong signals and one of which positionally overlaps linear epitopes in mustard and walnut allergenic 2S albumins. The overlapping cashew and walnut epitopes also share considerable homology. Conclusions: We conclude that this 2S albumin protein is a major allergen in cashew nut and demonstrates a possible basis for cross-reactivity with walnut 2S albumin.

KW - 2S albumin

KW - Ana o 3

KW - Cashew allergen

KW - Epitope mapping

KW - Food allergy

KW - Seed storage proteins

KW - Tree nut allergy

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