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Volume 272, Number 14, Issue of April 4, 1997 pp. 9539-9542
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Apoxin I, a Novel Apoptosis-inducing Factor with L-Amino Acid Oxidase Activity Purified from Western Diamondback Rattlesnake Venom

(Received for publication, August 13, 1996, and in revised form, January 21, 1997)

Shinichi Torii ^§ , Mikihiko Naito and Takashi Tsuruo ^¶

From the  Institute of Molecular and Cellular Biosciences, University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113, Japan, ^§ Janssen-Kyowa Co. Ltd., Takanawadai Daiichi-Seimai Building, Higashi-gotanda, Shinagawa-ku, Tokyo 141, Japan, and ^¶ Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Kami-ikebukuro, Toshima-ku, Tokyo 170, Japan

Venom of the western diamondback rattlesnake (Crotalus atrox) induces apoptosis in human umbilical vein endothelial cells, which could result in hemorrhage in tissues bitten by the snake. To identify the hemorrhagic factor, we purified a novel protein, apoxin I, from rattlesnake venom. Apoxin I induced apoptosis in human umbilical vein endothelial, human promyelocytic leukemia HL-60, human ovarian carcinoma A2780, and mouse endothelial KN-3 cells. Amino acid sequence analysis of the apoxin I showed close similarity to L-amino acid oxidase from the Malayan pit viper (Calloselasma rhodostoma). The purified apoxin I oxidized L-leucine but not D-leucine to produce H₂O₂. The apoxin I-induced apoptosis was inhibited by catalase, a H₂O₂ scavenger. These results indicate that the H₂O₂ produced by L-amino acid oxidation by apoxin I is involved in the apoxin I-induced apoptosis and in hemorrhage caused by rattlesnake venom.

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