Sarcoplasmic calcium-binding protein is an EF-hand–type protein identified as a new shrimp allergen Rosalía Ayuso, MD, PhD, Galina Grishina, MS, María.

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Presentation transcript:

Sarcoplasmic calcium-binding protein is an EF-hand–type protein identified as a new shrimp allergen Rosalía Ayuso, MD, PhD, Galina Grishina, MS, María Dolores Ibáñez, MD, PhD, Carlos Blanco, MD, PhD, Teresa Carrillo, MD, PhD, Ramon Bencharitiwong, MS, Silvia Sánchez, MD, Anna Nowak-Wegrzyn, MD, Hugh A. Sampson, MD Journal of Allergy and Clinical Immunology Volume 124, Issue 1, Pages 114-120 (July 2009) DOI: 10.1016/j.jaci.2009.04.016 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 IgE Immunoblot of a representative group of 14 patients to BS extract (A) and rSCP (B). P, Protein staining of total shrimp protein (A), rSCP (B). Lanes, Immunolabeling with 14 patients' sera; NC, negative controls (2 used in A, 3 in B). In red are marked subjects' sera used for ELISA experiments and in blue those used for the RBL assay. Levels of shrimp-specific IgE are noted in kUA/L. ND, Not Done. Journal of Allergy and Clinical Immunology 2009 124, 114-120DOI: (10.1016/j.jaci.2009.04.016) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Multiple sequence alignment of protein sequences of SCPs. Sequence identities of Lit v 4.0101 with Penaeus spp α chain (P02636; 93.8%), β chain (P02635; 80%), crayfish SCP (Pontascatus leptodactylus, ABB58783; Procambarus clarkii, P05946; 81-82%), scallop (Mizuhopecten yessoensis), SCP (P02637; 14%), and fruit fly SCP (Drosophila melanogaster NP_001015389 and NP_524381; 18% to 52%). Identical amino acids to L vannamei SCP are replaced by dashes. Peptides identified by MS/MS matching SCP α chain are marked in red, and those matching the SCP β chain in blue. Journal of Allergy and Clinical Immunology 2009 124, 114-120DOI: (10.1016/j.jaci.2009.04.016) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Immunoblot inhibition of IgE reactivity to rSCP with extracts from other arthropods and mollusks. P, Protein staining of rSCP. Lane 1, Pool without inhibitor. Lanes 2-9, pool preincubated with extract of D farinae (2), B germanica (3), boiled lobster (4), crab (5), squid (6), scallop (7), mussel (8), shrimp (9), and chickpea (10) as negative control. Journal of Allergy and Clinical Immunology 2009 124, 114-120DOI: (10.1016/j.jaci.2009.04.016) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 ELISA inhibition and mediator release from RBL cell line. ELISA inhibition of BS with serum from subject 2 (A) and subject 43 (B). Inhibitors: rSCP, rLit v 1, BS, and rCor a 9. Inhibitor concentrations ranged from 25 × 10-8 μg/mL (shown as 10-8) to 25 μg/mL total protein. β-Hexosaminidase release from RBL cells induced by rSCP, rLit v 1, and BS (maximal concentration, 1 μg/mL) is expressed as percentage of total possible release induced by Triton X-100. Cells were sensitized with serum pool of subjects 14, 47, and 48 (C). Journal of Allergy and Clinical Immunology 2009 124, 114-120DOI: (10.1016/j.jaci.2009.04.016) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions