Biochemical studies on proteinaceous toxins in the skin mucus and roe of the lamprey Lampetra japonica

七鳃鳗皮肤粘液和鱼子中蛋白质毒素的生化研究

• 批准号: 04660220
• 负责人: SHIOMI Kazuo
• 金额: $ 1.28万
• 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04660220/
• 关键词: Lamprey; Mucus toxin; Roe toxin; Proteinaceous toxins; Anguilled fish

1. Plroteinaceous toxins were newly found in the skin mucus and roe of the lamprey Lampetra japonica. One gram of the skin mucus and roe was evaluated to kill 70 and 215-800 mice with a body weight of 20g, respectively. Both toxins showed no hemagglutinating and hemolytic activities ; the roe toxin also lacked antimicrobial activity.2. The mucus toxin was very unstable and hence its purification was not achieved. On the other hand, the roe toxin was purified to an electrophoretically pure state by column chromatography on CM-cellulose and hydroxyapatite and FPLC on Mono S.The purified toxin exhibited high lethal activity upon i.v. injection into mice ; the LD_<50> was estimated to be 33 mug/kg. It was basic in nature and its molecular weight was determined to be 40,000 by SDS-PAGE in the absence of 2-mer captocthanal, Analysis by SDS-PAGE in the presence of the reducing reagent revealed that the toxin was comprised of two kinds of subunits with molecular weights of 30,000 and 10,000. The amino acid composition was characterized by low contents of sulfur-containing amino acids (half-Cys and Met).3. None of the 21 species of fish examined had toxins in their roes. in contrast, as many as 9 of 54 species of fish were found to contain toxins in the skin mucus. Toxins in 3 species of anguillid fish (Anguilla japonica, Anguilla anguilla and Muraenesox cinereus), which were highly toxic, were all acidic proteins with a molecular weight of about 400,000. They were also immunologically comparable to one another. It was noticeable that sphingosine and gangliosides showed inhibitory effects on the lethal activity of the A.japonica and A.anguilla toxins.

1.在日本七鳃鳗（Lampetra japonica）的皮肤粘液和鱼卵中发现了新的毒素。每克皮肤粘液和鱼卵分别杀死70只和215-800只体重为20 g的小鼠。两种毒素均无血凝和溶血活性，鱼卵毒素也无抗菌活性.粘液毒素非常不稳定，因此无法纯化。另一方面，通过在CM-纤维素和羟基磷灰石上的柱层析和在Mono S上的FPLC将鱼卵毒素纯化至药物纯状态。纯化的毒素在静脉注射到小鼠中时表现出高致死活性; LD<50>估计为33 μ g/kg。该毒素为碱性，在不加巯基乙酸的情况下，经SDS-PAGE测得其分子量为40，000。在加还原剂的情况下，经SDS-PAGE分析，该毒素由分子量为30，000和10，000的两种亚基组成。氨基酸组成以含硫氨基酸（半胱氨酸和蛋氨酸）含量低为特征.在21种被检测的鱼类中，没有一种在鱼卵中含有毒素。相比之下，在54种鱼类中，多达9种鱼类的皮肤粘液中含有毒素。日本鳗鲡（Anguillajaponica）、欧洲鳗鲡（Anguillaanguilla）和海鳗（Muraenesoxcinereus）等3种毒性较强的鳗类毒素均为酸性蛋白质，分子量约为40万。它们在免疫学上也彼此相当。鞘氨醇和神经节苷脂对日本血吸虫和欧洲鳗毒素的致死活性有明显的抑制作用。

项目成果

K.Shiomi: "Comparison of proteinaceous toxins in the skin mucus from three species of eels" Comp.Biochem.Physiol.107B. 389-394 (1994)

K.Shiomi：“三种鳗鱼皮肤粘液中蛋白质毒素的比较”Comp.Biochem.Physiol.107B。

K.Shiomi et al.: "Comparison of proteinaceous toxins in the skin mucus from three species of eels" Comp.Biochem.Physiol.107B. 389-394 (1994)

K.Shiomi 等人：“三种鳗鱼皮肤粘液中蛋白质毒素的比较”Comp.Biochem.Physiol.107B。