Isolation, cloning and characterization of the 2S albumin: a new allergen from hazelnut

A - Papers appearing in refereed journals

Garino, C., Zuidmeer, L., Marsh, J., Lovegrove, A., Morati, M., Versteeg, S., Schilte, P., Shewry, P. R., Arlorio, M. and Van Ree, R. 2010. Isolation, cloning and characterization of the 2S albumin: a new allergen from hazelnut. Molecular Nutrition & Food Research. 54 (9), pp. 1257-1265.

AuthorsGarino, C., Zuidmeer, L., Marsh, J., Lovegrove, A., Morati, M., Versteeg, S., Schilte, P., Shewry, P. R., Arlorio, M. and Van Ree, R.
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. 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. Results: N-terminal sequencing of a similar to 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 10aa internal peptide. The obtained clone (441 bp) encoded a 147aa 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. 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.

KeywordsFood Science & Technology
Year of Publication2010
JournalMolecular Nutrition & Food Research
Journal citation54 (9), pp. 1257-1265
Digital Object Identifier (DOI)doi:10.1002/mnfr.200900456
PubMed ID20373288
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
Regione Piemonte (Ricerca Sanitaria Finalizzata 2008)
Cariplo Foundation (Milan, Italy)
European Union - EU
Funder project or codeCentre for Crop Genetic Improvement (CGI)
ISSN16134125
PublisherWiley
Grant IDFOOD-CT-2005-514000

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