nsLTP and profilin are allergens in mustard seeds: cloning, sequencing and recombinant production of Sin a 3 and Sin a 4
Article first published online: 13 NOV 2009
© 2009 Blackwell Publishing Ltd
Clinical & Experimental Allergy
Volume 39, Issue 12, pages 1929–1936, December 2009
How to Cite
Sirvent, S., Palomares, O., Vereda, A., Villalba, M., Cuesta-Herranz, J. and Rodríguez, R. (2009), nsLTP and profilin are allergens in mustard seeds: cloning, sequencing and recombinant production of Sin a 3 and Sin a 4. Clinical & Experimental Allergy, 39: 1929–1936. doi: 10.1111/j.1365-2222.2009.03382.x
- Issue published online: 13 NOV 2009
- Article first published online: 13 NOV 2009
- Submitted 2 June 2009; revised 22 July 2009; accepted 8 September 2009.
- mustard allergy;
- Sin a 3;
- Sin a 4
Background Patients allergic to mustard are frequently sensitized to peach.
Objective To identify and analyse new yellow mustard allergens that could be involved in IgE cross-reactivity.
Methods Sera from mustard-allergic patients with symptoms to peach were studied. Mass spectrometry analyses provided sequences of IgE-reactive proteins. cDNAs encoding Sin a 3 and Sin a 4 were amplified by polymerase chain reaction, cloned and sequenced. The recombinant allergens were obtained in Pichia pastoris and Escherichia coli, respectively, and used for ELISA, immunoblotting and inhibition experiments. Sequence alignment was used to identify common IgE epitopes.
Results Sin a 3- and Sin a 4-specific cDNAs encode for mature proteins of 92 and 131 amino acids that belong to nsLTP and profilin protein families, respectively. Sin a 3 and Sin a 4 showed 54% and 80% identity with allergenic nsLTP from peach and profilin from melon, respectively. Both recombinant allergens were IgE-reactive in ELISA and immunoblotting. Peach pulp and peel, and melon extracts nearly abolished the IgE binding to recombinant Sin a 3 or recombinant Sin a 4 in immunoblotting.
Conclusion Sin a 3 (nsLTP) and Sin a 4 (profilin) were identified as new mustard allergens and showed IgE cross-reactivity with fruits such as peach or melon, respectively. The knowledge of these two allergens will contribute towards better understand with cross-reactivity between mustard and other plant food allergens, and their availability will provide physicians with useful tools for molecular diagnosis.