Biochemical studies on fish venoms

1. 6种海鱼（疣鱼、斑鱼、斑鱼、斑鱼、斑鱼、斑鱼）的毒液均表现出对小鼠的致死活性、对兔红细胞的特异溶血活性和透明质酸酶活性。市售石鱼抗蛇毒血清对石鱼毒的杀伤和溶血活性有较强的抑制作用。此外，石鱼抗毒血清也能中和其他五种毒液的致死性和溶血性活性，但程度较轻。这些结果表明，鱼类毒液在抗原性上具有相似性。毒毒液的致死因子不溶于蒸馏水，但可溶于高离子强度的溶剂。在TSK-gel G3000SW上，采用蒸馏水沉淀和高效液相色谱法，将其纯化为高效纯。纯化致死因子的LD_<50>为47马克杯/千克（小鼠静脉注射），并具有溶血和增加毛细血管通透性的活性。通过TSK-gel G3000SW凝胶过滤或SDS聚丙烯酰胺凝胶电泳测定其分子量约为90000，表明致死因子以单体蛋白的形式存在。疣状棘球蚴毒液在fsk -凝胶G3000SW上的行为；结果表明，在每种毒液中，同一因子同时具有致死性和溶血性活性。此外，不管毒液是什么，致死因子（溶血素）的分子量估计约为90000。透明质酸酶的分子量也被确定为60000-7000，与毒液无关。本研究检测的毒液的致死（和/或溶血）活性对冷冻和加热都非常不稳定。今后应建立一种稳定毒液的技术方法。

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.