Purification, biochemical, and immunological characterisation of a major food allergen

Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin

A. Bugajska-Schretter, H. Rumpold, S. Spitzauer, M. Grote, R. Reichelt, L. Vangelista, P. Valent, W. R. Sperr, A. Pastore, R. Valenta

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

Background - Almost 4% of the population suffer from food allergy which is an adverse reaction to food with an underlying immunological mechanism. Aims - To characterise one of the most frequent IgE defined food allergens, fish parvalbumin. Methods - Tissue and subcellular distribution of carp parvalbumin was analysed by immunogold electron microscopy and cell fractionation. Parvalbumin was purified to homogeneity, analysed by mass spectrometry and circular dichroism (CD) spectroscopy, and its allergenic activity was analysed by IgE binding and basophil histamine release tests. Results - The isoelectric point (pI) 4.7 form of carp parvalbumin, a three EF-hand calcium-binding protein, was purified to homogeneity. CD analysis revealed a remarkable stability and refolding capacity of calcium-bound parvalbumin. This may explain why parvalbumin, despite cooking and exposure to the gastrointestinal tract, can sensitise patients. Purified parvalbumin reacted with IgE of more than 95% of individuals allergic to fish, induced dose-dependent basophil histamine release and contained, on average, 83% of the IgE epitopes present in other fish species. Calcium depletion reduced the IgE binding capacity of parvalbumin which, according to CD analysis, may be due to conformation-dependent IgE recognition. Conclusions - Purified carp parvalbumin represents an important cross reactive food allergen. It can be used for in vitro and in vivo diagnosis of fish-induced food allergy. Our finding that the apo-form of parvalbumin had a greatly reduced IgE binding capacity indicates that this form may be a candidate for safe immunotherapy of fish-related food allergy.

Original languageEnglish
Pages (from-to)661-669
Number of pages9
JournalGut
Volume46
Issue number5
DOIs
Publication statusPublished - 2000
Externally publishedYes

Fingerprint

Parvalbumins
Carps
Allergens
Immunoglobulin E
Calcium
Food
Fishes
Food Hypersensitivity
Circular Dichroism
Basophils
Histamine Release
EF Hand Motifs
Cell Fractionation
Calcium-Binding Proteins
Isoelectric Point
Cooking
Tissue Distribution
Immunotherapy
Gastrointestinal Tract
Epitopes

Keywords

  • Antibodies
  • Circular dichroism
  • Epitopes
  • Food allergy
  • Immunochemistry
  • Parvalbumin

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Purification, biochemical, and immunological characterisation of a major food allergen : Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin. / Bugajska-Schretter, A.; Rumpold, H.; Spitzauer, S.; Grote, M.; Reichelt, R.; Vangelista, L.; Valent, P.; Sperr, W. R.; Pastore, A.; Valenta, R.

In: Gut, Vol. 46, No. 5, 2000, p. 661-669.

Research output: Contribution to journalArticle

Bugajska-Schretter, A, Rumpold, H, Spitzauer, S, Grote, M, Reichelt, R, Vangelista, L, Valent, P, Sperr, WR, Pastore, A & Valenta, R 2000, 'Purification, biochemical, and immunological characterisation of a major food allergen: Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin', Gut, vol. 46, no. 5, pp. 661-669. https://doi.org/10.1136/gut.46.5.661
Bugajska-Schretter, A. ; Rumpold, H. ; Spitzauer, S. ; Grote, M. ; Reichelt, R. ; Vangelista, L. ; Valent, P. ; Sperr, W. R. ; Pastore, A. ; Valenta, R. / Purification, biochemical, and immunological characterisation of a major food allergen : Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin. In: Gut. 2000 ; Vol. 46, No. 5. pp. 661-669.
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abstract = "Background - Almost 4{\%} of the population suffer from food allergy which is an adverse reaction to food with an underlying immunological mechanism. Aims - To characterise one of the most frequent IgE defined food allergens, fish parvalbumin. Methods - Tissue and subcellular distribution of carp parvalbumin was analysed by immunogold electron microscopy and cell fractionation. Parvalbumin was purified to homogeneity, analysed by mass spectrometry and circular dichroism (CD) spectroscopy, and its allergenic activity was analysed by IgE binding and basophil histamine release tests. Results - The isoelectric point (pI) 4.7 form of carp parvalbumin, a three EF-hand calcium-binding protein, was purified to homogeneity. CD analysis revealed a remarkable stability and refolding capacity of calcium-bound parvalbumin. This may explain why parvalbumin, despite cooking and exposure to the gastrointestinal tract, can sensitise patients. Purified parvalbumin reacted with IgE of more than 95{\%} of individuals allergic to fish, induced dose-dependent basophil histamine release and contained, on average, 83{\%} of the IgE epitopes present in other fish species. Calcium depletion reduced the IgE binding capacity of parvalbumin which, according to CD analysis, may be due to conformation-dependent IgE recognition. Conclusions - Purified carp parvalbumin represents an important cross reactive food allergen. It can be used for in vitro and in vivo diagnosis of fish-induced food allergy. Our finding that the apo-form of parvalbumin had a greatly reduced IgE binding capacity indicates that this form may be a candidate for safe immunotherapy of fish-related food allergy.",
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AU - Spitzauer, S.

AU - Grote, M.

AU - Reichelt, R.

AU - Vangelista, L.

AU - Valent, P.

AU - Sperr, W. R.

AU - Pastore, A.

AU - Valenta, R.

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AB - Background - Almost 4% of the population suffer from food allergy which is an adverse reaction to food with an underlying immunological mechanism. Aims - To characterise one of the most frequent IgE defined food allergens, fish parvalbumin. Methods - Tissue and subcellular distribution of carp parvalbumin was analysed by immunogold electron microscopy and cell fractionation. Parvalbumin was purified to homogeneity, analysed by mass spectrometry and circular dichroism (CD) spectroscopy, and its allergenic activity was analysed by IgE binding and basophil histamine release tests. Results - The isoelectric point (pI) 4.7 form of carp parvalbumin, a three EF-hand calcium-binding protein, was purified to homogeneity. CD analysis revealed a remarkable stability and refolding capacity of calcium-bound parvalbumin. This may explain why parvalbumin, despite cooking and exposure to the gastrointestinal tract, can sensitise patients. Purified parvalbumin reacted with IgE of more than 95% of individuals allergic to fish, induced dose-dependent basophil histamine release and contained, on average, 83% of the IgE epitopes present in other fish species. Calcium depletion reduced the IgE binding capacity of parvalbumin which, according to CD analysis, may be due to conformation-dependent IgE recognition. Conclusions - Purified carp parvalbumin represents an important cross reactive food allergen. It can be used for in vitro and in vivo diagnosis of fish-induced food allergy. Our finding that the apo-form of parvalbumin had a greatly reduced IgE binding capacity indicates that this form may be a candidate for safe immunotherapy of fish-related food allergy.

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