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Isolation, cloning, and characterization of the 2S albumin: a new allergen from hazelnut.
- Source :
-
Molecular nutrition & food research [Mol Nutr Food Res] 2010 Sep; Vol. 54 (9), pp. 1257-65. - Publication Year :
- 2010
-
Abstract
- Scope: 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity.<br />Methods: 2S albumin from hazelnut extract was purified using size exclusion chromatography and RP-HPLC. After N-terminal sequencing, degenerated and poly-d(T) primers were used to clone the 2S albumin sequence from hazelnut cDNA. After expression in Escherichia coli and affinity purification, IgE reactivity was evaluated by Immunoblot/ImmunoCAP (inhibition) analyses using sera of nut-allergic patients.<br />Results: N-terminal sequencing of a approximately 10 kDa peak from size exclusion chromatography/RP-HPLC gave two sequences highly homologous to pecan 2S albumin, an 11 amino acid (aa) N-terminal and a 10 aa internal peptide. The obtained clone (441 bp) encoded a 147 aa hazelnut 2S albumin consisting of a putative signal peptide (22 aa), a linker peptide (20 aa), and the mature protein sequence (105 aa). The latter was successfully expressed in E. coli. Both recombinant and natural 2S albumin demonstrated similar IgE reactivity in Immunoblot/ImmunoCAP (inhibition) analyses.<br />Conclusion: We confirmed the postulated role of hazelnut 2S albumin as an allergen. The availability of recombinant molecules will allow establishing the importance of hazelnut 2S albumin for hazelnut allergy.
- Subjects :
- Adolescent
Adult
Aged
Amino Acid Sequence
Corylus metabolism
Female
Humans
Immunoglobulin E immunology
Immunoglobulin E metabolism
Male
Molecular Sequence Data
Nut Hypersensitivity blood
Nuts metabolism
Plant Extracts metabolism
Protein Precursors chemistry
Protein Precursors genetics
Protein Precursors immunology
Protein Precursors isolation & purification
Protein Sorting Signals
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins immunology
Recombinant Fusion Proteins isolation & purification
Sequence Alignment
Sequence Homology
Young Adult
2S Albumins, Plant chemistry
2S Albumins, Plant genetics
2S Albumins, Plant immunology
2S Albumins, Plant isolation & purification
Allergens chemistry
Allergens genetics
Allergens immunology
Allergens isolation & purification
Corylus immunology
Nut Hypersensitivity immunology
Nuts immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1613-4133
- Volume :
- 54
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Molecular nutrition & food research
- Publication Type :
- Academic Journal
- Accession number :
- 20373288
- Full Text :
- https://doi.org/10.1002/mnfr.200900456