Biochemical studies on fish venoms

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1. All venoms from six species of marine fish (Synanceja verrucosa, lminicus japonicus, Pterois lunulatas, P. volitans, P. antennatat, Dendrochirus zebra) exhibited lethal activity against mice, hemolytic activity specific for rabbit erythrocytes and hyaluronidase activity.2. The commercial stonefish antivenom raised against the Synanceia trachynis venom strongly inhibited the lethal and hemolytic activities of S. verrucosa venom. In addition, the lethal and hemolytic activities of the other five venoms were also neutralized by the stonefish antivenom, though to a lesser extent. These results suggested that fish venoms are antigenically similar to each other.3. The lethal factor in the S. verrucosa venom was found to be insoluble in distilled water but soluble in solvents with high ionic strength. It was purified to an efectrophoretically pure state by a very simple method comprising of precipitation in distilled water and HPLC on TSK-gel G3000SW. The purified lethal factor had a LD_<50> of 47 mug/kg (i.v. injection into mice) and also had hemolytic and capillary permeability-increasing activities. The molecular weight was determined to be about 90000 by either gel filtration on TSK-gel G3000SW or SDS polyacrylamide gel electrophoresis, suggesting that the lethal factor exists as a monomeric protein.4. Behaviors on FSK-gel G3000SW of the venoms from S. verrucosa, 1. japonicus, P. lunulata and P. volitans revealed that in each venom the same factor has both lethal and hemolytic activities. Furthermore, the molecular weight of the lethal factor (hemolysin) was estimated to be about 90000, irrespective of the venoms. The molecular weight of the hyaluronidase was also determined to be 60000-7000, irrespective of the venoms.5. The lethal (and/or hemolytic) activity of the venoms examined in this study were all very unstable to freezing and heating. In future a skillful method for stabilizing venoms should be established.