Molecular Cloning and Functional Analysis of Apoxin I, a Snake Venom-Derived Apoptosis-Inducing Factor with<scp>l</scp>-Amino Acid Oxidase Activity
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- Shinichi Torii
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Kazuhiko Yamane
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Tetsuo Mashima
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Naomi Haga
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Kazuo Yamamoto
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Jay W. Fox
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Mikihiko Naito
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
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- Takashi Tsuruo
- Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Janssen-Kyowa CO. Ltd., Takanawadai Daiichi-Seimei Bldg, Higashi-Gotanda, Shinagawa-ku, Tokyo 141, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-Ikebukuro, Toshima-ku, Tokyo 170, Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113−0033, and Department of Microbiology, University of Virginia,...
Description
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 F1G1 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 posttranslational modification, such as N-glycosylation, are required.
Journal
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- Biochemistry
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Biochemistry 39 (12), 3197-3205, 2000-03-01
American Chemical Society (ACS)
