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Biochemical studies on fish venoms

鱼毒的生化研究

基本信息

批准号：
63560202
负责人：
SHIOMI Kazuo
金额：
$ 1.41万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560202/
关键词：
Fish Stonefish Venom Lethal activity Hemolytic activity Hyaluronidase Stonefish antivenom Purification 生物活性

项目摘要

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.

1. 6种海洋鱼类（多疣梭子鱼、小梭子鱼、新月狐蝠、伏翼梭子鱼、触角梭子鱼、斑马树手鱼）的蛇毒均具有对小鼠的致死活性、对兔红细胞的溶血活性和透明质酸酶活性.用抗粗纹银汉鱼毒的抗石鱼血清对粗纹银汉鱼毒的致死和溶血活性有明显的抑制作用。疣状毒液此外，其他五种毒液的致死和溶血活性也被石鱼抗蛇毒血清中和，尽管程度较低。这些结果表明，鱼类毒液具有相似的抗原性. S.发现疣状毒液不溶于蒸馏水，但溶于具有高离子强度的溶剂。通过非常简单的方法将其纯化至电泳纯状态，所述方法包括在蒸馏水中沉淀和在TSK-凝胶G3000 SW上的HPLC。纯化的致死因子的LD为<50>47 μ g/kg（静脉注射到小鼠体内），并且还具有溶血和毛细血管通透性增加活性。经TSK-gel G3000 SW凝胶过滤或SDS聚丙烯酰胺凝胶电泳测定，其分子量约为90000，表明该致死因子以单体蛋白形式存在.在FSK-gel G3000 SW上的行为。verrucosa，1. japonicus、P. lunulata和P. volitans的研究表明，在每种毒液中，相同的因子都具有致死和溶血活性。此外，致死因子（溶血素）的分子量估计为约90000，无论毒液。透明质酸酶的分子量也被确定为60000-7000，与毒液无关。本研究中所检测的毒液的致死（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巧妙方法。