2002 Fiscal Year Final Research Report Summary
Study on tropomyosin, a common and major allergen in Crustacea
| Project/Area Number |
13680157
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
食生活
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| Research Institution | Okayama Prefectural University |
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Principal Investigator |
KIMOTO Masumi Okayama Pref. Univ. Fac. Health and Welfare Science, Professor, 保健福祉学部, 教授 (40108866)
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| Co-Investigator(Kenkyū-buntansha) |
TERAMOTO Ai Okayama Pref. Univ. Fac. Health and Welfare Science, Assistant Researcher, 保健福祉学部, 助手 (50275369)
FUCHIGAMI Michiko Okayama Pref. Univ. Fac. Health and Welfare Science, Professor, 保健福祉学部, 教授 (60079241)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Shrimp Allergen / Monoclonal antibody / Sandwich ELISA / Tropomyosin / cDNA cloning / Epitope / Immunoblot / IgE antibody |
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| Research Abstract |
Recently, in Japan, many patients are suffering from allergy eliciting by ingestion of shrimp. Shrimp is an important foodstuff of the animal protein in view of its consumption quantity in this country. In order to elucidate the allergenicity of shrimp, we screened the allergens in it using the sera of shrimp-sensitive patients with atopic dermatitis and found only one protein, tropomyosin. This is a protein component, which form the muscle proteins in vertebrates and invertebrates, and is reported as a common allergic protein in seafood as well as Cepharopoda, Mollusca, Crustacea. In the present study, we attempted to purify tropomyosin (Pen j 1) from Penaceus japonicus, and prepared the monoclonal and polyclonal antibodies raised against the purified tropomyosin We investigated hypoallergenicity during cooking process by means of a sandwich enzyme-linked immunosorbent assay (ELISA) system established using the monoclonal and polyclonal antibodies obtained. Most of tropomyosin was extracted in the boiling water, suggesting that boiling process in cooking is effective in hypoallergenicity of shrimp.
Furthermore, we investigated the cloning of cDNA encoding the allergen to reveal information concerning its complete primary structure and allergenicity. The cDNA clone was cloned by means of 5'- and 3'-RACE method using mRNA prepared from fresh shrimp as a template and encoded a protein consist of 284 amino acids. Further, a various of recombinant peptides were expressed in E. coli to elucidate the epitopes, the binding region with IgE in patients' sera. The result of epitope mapping showed that the epitopes occurred and were divided into several segments over a very wide region on the polypeptide chain of tropomyosin.
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Research Products
(2 results)
