Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen - 13/08/11
, Fumihiro Arakawa, MS b, , Hiroshi Akiyama, PhD c, ⁎ 
, Yasuo Yoshioka, PhD c, Yoshiki Hayashi, PhD c, Shinobu Sakai, PhD c, Sumio Ito, MS b, Yuko Yamakawa, MD, PhD d, Satoru Ohgiya, PhD e, f, Zenro Ikezawa, MD, PhD d, Reiko Teshima, PhD cReprint requests: Hiroshi Akiyama, PhD, Section Chief, Division of Novel Foods and Immunochemistry, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.Abstract |
Background |
Carmine is a natural red pigment obtained from dried gravid female cochineal insects (Dactylopius coccus or Coccus cacti). There have been several reports of allergies to carmine, but the major allergens responsible have not been identified.
Objective |
To identify the major allergenic proteins in cochineal.
Methods |
Immunoblots of purified cochineal extract were probed with sera from 3 patients with allergy. Partial amino acid sequences were determined for the proteins bound by IgE, and the corresponding cDNA, containing a complete coding region, was cloned by 5′ and 3′ rapid cDNA extension and PCR. The recombinant protein was expressed in yeast and subjected to immunoblotting.
Results |
We identified a full-length cDNA encoding a protein, which we named CC38K, with 335 amino acids and a molecular mass calculated as 38 kd. This amino acid sequence included all the partial amino acid sequences obtained from the purified proteins identified by IgE from patients with allergy. Recombinant CC38K protein was recognized by patients’ sera, indicating that this is a major allergen present in carmine. The CC38K sequence showed homology to phospholipases.
Conclusion |
We have, for the first time, identified the major allergen in cochineal extract. This protein may be a phospholipase or related enzyme, both of which are known to be allergens in other insects.
Le texte complet de cet article est disponible en PDF.Key words : Cochineal, allergen, cDNA, IgE, food allergy, carmine, immunoblotting, cloning, recombinant
Abbreviations used : C1, C22, CC38K, 38K cDNA, PLA1, RACE
Plan
| Supported in part by grants from the Ministry of Health, Labor and Welfare of Japan and the Japan Food Chemical Research Foundation. Supported by a training program for building research capacity in the field of epidemiology and public health science in the National Institute of Public Health, Wako, Japan. |
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| Disclosure of potential conflict of interest: S. Ohgiya has received research support from Mitsui Engineering Corp and the Ministry of Industry, Trade, and Economy (Japan). The rest of the authors have declared that they have no conflict of interest. |
Vol 123 - N° 5
P. 1157 - mai 2009 Retour au numéro
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