| Project/Area Number |
14360045
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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 |
応用微生物学・応用生物化学
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| Research Institution | Tohoku University |
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Principal Investigator |
OGAWA Tomohisa Tohoku University, Graduate School of Life Sciences, Associate Professor, 大学院・生命科学研究科, 助教授 (80240901)
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| Co-Investigator(Kenkyū-buntansha) |
MURAMOTO Koji Tohoku University, Graduate School of Life Sciences, Professor, 大学院・生命科学研究科, 教授 (90157800)
NIISHIDA Yoshihiro Nagoya University, Graduate School of Engineerings, Associate Professor, 工学研究科, 助教授 (80183896)
SHIRAI Tsuyoshi Tohoku University, Biomolecular Engineering Research Institute, Chief scientist, 情報解析研究部門, 主任研究員 (00262890)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,600,000 (Direct Cost: ¥14,600,000)
Fiscal Year 2004: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2003: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥8,500,000 (Direct Cost: ¥8,500,000)
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| Keywords | Galectin / Congerin / Apoptosis / Carbohydrate recognition / Lectin / 糖鎖 / グライコーム / T細胞 / 加速進化 / グライコ-ム / 精密糖鎖認識 |
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| Research Abstract |
Two galectins termed Congerin I and Congerin II (Con1 and Con2) have been identified from skin mucus of conger eel. They showed 48% sequence identity with different carbohydrate-binding specificity. To analysis the detailed structure and carbohydrate binding ability relationships of congeries, in this study, reciprocal substituted mutants between Con1 and Con2 at the amino acid residues located in peripheral carbohydrate-binding clefts were prepared by site-direct mutagenesis. Analyses of their binding-specificity against 34 kinds of pyridylaminated oligosaccharides using frontal affinity chromatography system revealed that they had unique sugar specificities. For examples, it was found that V68N, Q71E, Q72T, E74Q and T107N mutations of Congerins affected on the strong carbohydrate binding activity. On the other hand, Congerins have already known had strong inducing activity for apoptotic cell death of human T cell lines, Jurkat cells. In this study, the cytotoxic activities of Congerin mutants were assayed using Jurkat cells to evaluate their biological activities. Comparison of ED50 of each mutant showed clearly correlations between apoptotic activities and sugar binding activities of Congerin mutants, suggesting that the specific sugar recognition was important for the apoptotic activity. Thus, it has been elucidated that the Congerin mutants obtained unique sugar binding profiles are applicable as tools for analyzing the structure and activities of unique carbohydrates.
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