Sarcoplasmic calcium-binding protein is an EF-hand–type protein identified as a new shrimp allergen - 15/08/11
, Galina Grishina, MS a, María Dolores Ibáñez, MD, PhD b, Carlos Blanco, MD, PhD c, Teresa Carrillo, MD, PhD d, Ramon Bencharitiwong, MS a, Silvia Sánchez, MD e, Anna Nowak-Wegrzyn, MD a, Hugh A. Sampson, MD a
Reprint requests: Rosalía Ayuso, MD, PhD, Mt Sinai School of Medicine, One Gustave L. Levy Place, Box 1198, New York, NY 10029.Abstract |
Background |
Shellfish allergy is a long-lasting disorder usually persisting throughout life. Despite its high prevalence, there is limited information about allergenic shrimp proteins.
Objective |
Characterization of shrimp allergens.
Methods |
Fifty-two adults and children with a history of immediate allergic reactions to shrimp and elevated serum IgE to shrimp were selected for this study. Tryptic digests from a 20-kd IgE-binding protein were analyzed by LC-MS/MS, identifying the protein as a sarcoplasmic-calcium-binding protein. cDNA encoding sarcoplasmic calcium-binding protein (SCP) from a shrimp cDNA library (Litopenaeus vannamei) was amplified by PCR, cloned into an expression vector, and sequenced. Recombinant SCP was tested with patients’ sera. ELISA inhibition experiments determined the fraction of total shrimp IgE recognizing SCP. A functional assay with a rat basophilic leukemia cell line was used to determine the capacity for mediator release induced by SCP.
Results |
Immunoblotting demonstrated IgE binding by 31 of 52 (59.6%) of the sera to a 20-kd shrimp protein. The protein was identified as a SCP. Amplified cDNA encoding SCP was isolated and sequenced. Open reading frame translation provided the complete amino acid sequence of shrimp SCP. Recombinant SCP was recognized by serum IgE from 20 of 52 (38.4%) subjects, of whom 17 of 20 (85%) were children. ELISA inhibition of pooled sera IgE reactivity to BS extract using recombinant SCP was significant (as high as 79%). For some subjects, mediator release induced by recombinant SCP was higher than that induced by recombinant tropomyosin.
Conclusion |
We have identified and cloned a new shrimp allergen, Lit v 4.0101, an SCP, which appears to be of particular importance in the pediatric population.
Le texte complet de cet article est disponible en PDF.Key words : Allergen, Crustacea, shellfish, shrimp, recombinant, Lit v 4, sarcoplasmic calcium-binding protein
Abbreviations used : BS, MLC, LC-MS/MS, MW, r, RBL, SCP
Plan
| Supported by the Food Allergy Initiative. A.N.-W. is supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, grant no. AI 059318. |
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| Disclosure of potential conflict of interest: R. Ayuso has performed grant research for the Food Allergy Initiative. A. Nowak-Wegrzyn is on the Safety Board for Schering-Plough, has received research support from Novartis Vaccines, and is Vice President and Treasurer for the New York Allergy and Asthma Society. H. A. Sampson is a consultant and 4% shareholder for Allertein Pharmaceuticals, LLC; is on the advisory board for Schering-Plough; has received research support from the Food Allergy Initiative and the National Institute of Allergy and Infectious Diseases, National Institutes of Health; and is President of the American Academy of Allergy, Asthma & Immunology. The rest of the authors have declared that they have no conflict of interest. |
Vol 124 - N° 1
P. 114-120 - juillet 2009 Retour au numéro
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