2000 Fiscal Year Final Research Report Summary
Molecular function and biosynthesis of apoxin I, an apoptosis inducing factor derived from snake venom.
| Project/Area Number |
11672159
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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 |
Biological pharmacy
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| Research Institution | Institute of Molecular and Cellular Biosciences, The University of Tokyo |
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Principal Investigator |
NAITO Mikihiko Associate Professor, Institute of Molecular and Cellular Biosciences, The University of Tokyo, 分子細胞生物学研究所, 助教授 (00198011)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Apoxin / L-amino acid oxidase / snake venom / apoptosis |
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| Research Abstract |
We previously purified apoxin I, an apoptosis-inducing factor with L-amino acid oxidase (LAO) activity, from Western diamondback rattlesnake venom. To determine the primary structure of apoxin I, we cloned its cDNA.The amino acid sequence showed that apoxin I has an FAD binding domain and shares homology with L-amino acid oxidase (LAO) from Neurospora crassa, human monoamine oxidase B and mouse interleukin 4-induced Fig1 protein. The full-length apoxin I has an N-terminal signal sequence that is processed in mature apoxin I in venom. When the apoxin I gene was transfected into human 293T cells, the recombinant protein was expressed in the cells, and a significant amount of apoxin I was secreted into the medium. The secreted recombinant apoxin I protein showed LAO and apoptosis-inducing activity, but the recombinant protein in the cells did not, suggesting that maturation and secretion of the apoxin I protein is needed for its activity. Treating the transfected cells with tunicamycin inhibited the secretion and LAO activity of the recombinant apoxin I.In addition, deleting the amino-terminal region flanking the signal sequence, the FAD binding domain and the carboxy-terminal region abolished the secretion and LAO activity of the recombinant proteins. These results indicate that in order for apoxin I to become active these regions and post-translational modification, such as N-glycosylation, are required.
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[Publications] Torii, S., Yamane, K., Mashima, T., Haga, N., Yamamoto, K., Fox, J.W., Naito, M., and Tsuruo, T.: "Molecular cloning and functional analysis of apoxin I, a snake venom-derived apoptosis-inducing factor with L-amino acid oxidase activity."Biochemistry. 39. 3197-3205 (2000)
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