| Project/Area Number |
17380124
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Fisheries chemistry
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| Research Institution | Tokyo University of Marine Science and Technology |
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Principal Investigator |
SHIOMI Kazuo Tokyo University of Marine Science and Technology, Faculty of Marine Science, Professor, 海洋科学部, 教授 (90111690)
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| Co-Investigator(Kenkyū-buntansha) |
USHIO Hideki Tokyo University of Marine Science and Technology, Faculty of Marine Science, Associate Professor, 海洋科学部, 助教授 (50251682)
ISHIZAKI Shoichiro Tokyo University of Marine Science and Technology, Faculty of Marine Science, Associate Professor, 海洋科学部, 助教授 (40251681)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2006: ¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 2005: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | seafood / allergen / parvalbumin / tropomyosin / collagen / cDNA cloning / antibody / IgE-binding epitope / モノクローナル抗体 |
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| Research Abstract |
1.Amino acid sequences of tropomyosins (TMs) from four species of crustaceans and ten species of shellfish were elucidated by cDNA cloning. The amino acid sequence of acorn barnacle TM was significantly divergent from those of crustacean TMs, while TMs from the other three species of crustaceans showed high sequence identity (>90% identity) with the known crustacean TMs. On the other hand, the amino acid sequences of shellfish TMs were considerably different from those of crustacean and cephalopod TMs and were also conserved only within the same family (or order).
2.Mackerel parvalbumin (PA) exhibited a significantly reduced IgE-binding ability upon Ca^<2+>-depletion. Moreover, the IgE-binding ability of PA was markedly reduced by replacements of Asp-51 and Asp-90, which are involved in the Ca^<2+>-binding, by Ala. These results led us to assume that the IgE-binding epitopes of mackerel PA are largely associated with the solid conformation formed by Ca^<2+>-chelating.
3.ELISA using synthetic peptides revealed that the major linear-type IgE-binding epitope of mackerel PA is contained in the region 21-40, that of rainbow trout collagen α2 chain in the region 941-968 and those of Japanese flying squid TM in the regions 113-132, 185-204 and 265-284.
4.A 20 kDa allergen was newly found in black tiger shrimp and identified as a sarcoplasmic calcium-binding protein by the determined partial amino acid sequence.
5.An ECL-western blotting method with high sensitivity was developed for the detection of PA. Application of this method to fish collagen products showed the contamination of trace amounts of PA.
6.Experiments using five model peptides (10-12 amino acids long) confirmed an effective method for the production of antibodies against peptidic antigens. This method was assumed to be applicable to the production of specific antibodies recognizing IgE-binding epitopes of fish allergens.
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