1988 Fiscal Year Final Research Report Summary
Analysis of diphtheria toxin receptor by molecular cloning.
| Project/Area Number |
62570189
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Kurume University (1988)
Osaka University (1987) |
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Principal Investigator |
MEKADA Eisuke Kurume University, 分子生命科学研究所, 教授 (20135742)
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| Co-Investigator(Kenkyū-buntansha) |
KANEDA Yasufumi Institute for Molecular and Cellular Biology, Osaka Univ., 細胞工学センター, 助手 (10177537)
UCHIDA Tsuyoshi Institute for Molecular and Cellular Biology, Osaka Univ., 細胞工学センター, 教授 (40029781)
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| Project Period (FY) |
1987 – 1988
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| Keywords | Diphtheria toxin / Receptor / Monoclonal Antibody / エンドサイドーシス |
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| Research Abstract |
The aim of this project is to understand the entry mechanism of diphtheria toxin into cytosol. The strategy of this project is as follows: (1)Identification of diphtheria toxin receptor in membrane fractions of cultured cells. (2)Purification of the receptor. (3)Isolation of monoclonal antibodies against diphtheria toxin receptor. (4)cDNA cloning of the receptor using monoclonal antobodies.
In 1987, we identified diphtheria toxin receptor as 14.5Kd protein in lysate of Vero cell membrane using CRM197 protein, a mutant protein of diphtheria toxin. Then the receptor was purified by a 10,000-folds by sequential chromatography. In this year, we tried to isolate hybridomas producing antibodies directed to diph-theria toxin receptor. Several clones were isolated. An antibody produced from one of these clones inhibited the binding of diphtheria toxin to the receptor. This antibody specefically precipitates the 14.5Kd protein in the fraction of partially purified receptor. Now we tried to isolate cDNA of the receptor using these monoclonal antibodies.
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