Structure and Function of Proteinaceous Toxins from Marine Animals

海洋动物蛋白质毒素的结构和功能

• 批准号: 12045267
• 负责人: NAGAI Hiroshi
• 金额: $ 8.19万
• 依托单位: Tokyo University of Marine Science and Technology (Tokyo University of Fisheries) (2001-2003)Suntory Institute for Bioorganic Research (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12045267/
• 关键词: protein; venom; toxin; jellyfish; sea anemone; sting; 刺胞; 生理活性; ハブクラゲ; ミズクラゲ; タンパク質毒素; 溶血; Carybdea alata; アンドンクラゲ

Two liable potent hemolytic toxins from the tentacles of the stinging box jellyfish (sea wasp) Carybdea rastoni were isolated and sequenced, C. rastoni toxin-A and -B (CrTX-A and -B). CrTX-A and -B which have molecular masses of 43 and 46 kDa (calculated from (SDS-PAGE), respectively. The full-length cDNA (1600bp) which encodes CrTXs was sequenced. The deduced amino acid sequence of CrTXs, composed of 450 amino acids, showed no significant homology with any known protein. This is the first complete sequence of a jellyfish proteinous toxin to be reported. The experimental results showed CrTX-A was a toxin responsible for the cutaneous inflammation observed in humans inflicted by C. rastoni's sting. The box jellyfish (Sea Wasp) Carybdea alata (Cubozoa) is distributed widely in the tropics. We successfully isolated C. alata toxin-A (CaTX-A, 43kDa) and -B (CaTX-B, 45kDa) for the first time from the tentacle of C. alata collected at the Hawaiian shore. Furthermore, we sequenced the cDNA enc … More oding CaTX-A. We successftlly isolated Chiropsalmus quadrigatus toxin-A (CqTX-A, 44 kDa), a major proteinaceous toxin, for the frst time, from the deadly box jellyfish C. quadrigatus. We sequenced the cDNA encoding CqTX-A. The deduced amino acid sequence of CqTX-A (462 amino acids) showed 25.2% and 21.6% sequence similarity to Carybdea rastoni toxins (CrTXs) and Carybdea alata toxin-A (CrTX-A). The box jellyfish toxins represent a novel bioactive protein family.The Okinawan sea anemone Phyllodiscus semoni is known to cause cases of severe stinging. We isolated Phyllodiscus semoni toxin 20A (PsTX-20A), a hemolytic and lethal polypeptide (20 kDa), from this species for the first time as a minor toxin. Furthermore, we sequenced the cDNA encoding PsTX-20A. The deduced amino acid sequence of PsTX-20A showed that this toxin was a new member of the actinoporin family, which consists of several cytolytic polypeptides originating from sea anemones. We isolated Phyllodiscus semoni toxins 60A and 60B (PsTX-60A and PsTX-60B ; ca. 60 kDa) as the major toxins of this species. We sequenced the cDNA encoding PsTX-60A. We also isolated Actineria villosa toxin 60A (AvTX-60A, 60 kDa) as the major toxin from this venomous sea anemone. Furthermore, the cDNA encoding AvTX-60A was sequenced. The deduced amino acid sequence of AvTX-60A showed similarity with PsTX-60A. It was demonstrated that these sea anemone toxins were new members of a membrane-attack complex/ perforin (MACPF) family. These are the first examples of MACPF type proteins as the toxins for prey acquisition or repelling predators in nature.Our study showed the venomous cnidarians are rich sources of novel bioactive proteins. Further chemical and pharmacological study on the cnidarian toxins will lead to discover new tools for bio-medical study, proteinaceous drugs which have new mode of action, and develop of new remedy methods for these animal's stinging cases. Less

从刺盒水母(海蜂)触角中分离到两种敏感的强效溶血毒素，并对其进行了序列测定，它们是刺盒水母毒素A和B(CRTX-A和-B)。CRTX-A和CRTX-B的相对分子质量分别为43 kDa和46 kDa(由(SDS-PAGE)计算)。对编码CrTXs的全长cDNA1600bp进行了测序。其推导的氨基酸序列由450个氨基酸组成，与已知的任何蛋白质没有明显的同源性。这是首次报道的水母蛋白毒素的完整序列。实验结果表明，CRTX-A是一种毒素，可导致人体被拉斯托尼弧菌叮咬引起的皮肤炎。箱水母(海蜂)是一种广泛分布于热带地区的水母。本研究首次从夏威夷海岸采集的银鱼触须中分离到银鱼毒素-A(CATx-A，43 kDa)和毒素-B(CATx-B，45 kDa)。此外，我们还对cdna enc…进行了测序。更多ODING CATx-A我们首次成功地从致死的箱形水母中分离到一种主要的蛋白质毒素-A(CqTX-A)。我们对编码CqTX-A的基因进行了测序。CqTX-A全长462个氨基酸，与鸡冠鱼毒素(CrTXs)和翼鱼毒素-A(CRTX-A)的同源性分别为25.2%和21.6%。盒状水母毒素代表了一个新的生物活性蛋白家族。冲绳海葵叶状海葵已知会引起严重的叮咬。本研究首次从该物种中分离到一种溶血致死的多肽--叶盘藻毒素20A(PSTX-20A)，作为次要毒素。此外，我们还对pstx-20A的编码基因进行了测序。PSTX-20A的氨基酸序列分析表明，该毒素是放线菌素家族的一个新成员，由来源于海葵的几个细胞溶解多肽组成。我们分离到叶盘藻毒素60A和60B(pstx-60A和pstx-60B，约60 kDa)作为该物种的主要毒素。我们对编码pstx-60A的基因进行了测序。我们还从这种有毒的海葵中分离出了主要毒素--簇毛放线菌毒素60A(AvTX-60A，60 kDa)。此外，还对AvTX-60A的编码基因进行了测序。AvTX-60A的氨基酸序列与pSTX-60A相似。这些海葵毒素被证明是膜攻击复合体/穿孔素(MACPF)家族的新成员。这是在自然界中首次发现MACPF类蛋白作为猎物获取或驱赶捕食者的毒素。我们的研究表明，有毒的线虫是新的生物活性蛋白的丰富来源。对其进行进一步的化学和药理研究，将为生物医学研究发现新的工具和具有新的作用机制的蛋白质类药物，并为这些动物的叮咬病例开发新的治疗方法。较少

项目成果

Emikko Ito, Hiroshi Nagai: "Bleeding from the small intestine caused by aplysiatoxin, the causative agent of the red alga Gracilaria coronopifolia poisoning."Toxicon. 38. 123-132 (2000)

Emikko Ito、Hiroshi Nagai：“海兔毒素引起的小肠出血，海兔毒素是红藻江蓠中毒的病原体。”Toxicon。

Ukai et al.: "Polyhydroxysteroids and Saponins of Asterino Pectinifera"Biosci. Biotech. Biochem.. 66. 913-915 (2002)

Ukai 等人：“海星的多羟基类固醇和皂苷”Biosci。

永井宏史: "初めて明らかにされたクラゲ毒の化学的性状(アンドンクラゲから得られたタンパク質毒素)"バイオサインエスとインダストリー. 58. 43-44 (2000)

Hiroshi Nagai：“首次揭示水母毒素的化学性质（从安灯水母中获得的蛋白质毒素）”《生物科学与工业》58. 43-44 (2000)。

永井宏史: "立方クラゲ類の刺胞毒"日本水産学会誌. 69. 827-828 (2003)

Hiroshi Nagai：“立方水母的刺丝囊毒”日本水产学会杂志 69. 827-828 (2003)。

Hiroshi Nagai, Kyoko Takuwa, Masahiro Nakao, Emiko Ito, Masami Miyake, Masatoshi Noda, Terumi Nakajima: "Novel Proteinaceous Toxins from the Box Jellyfish (Sea Wasp) Carybdea rastoni"Biochem. Biophys. Res. Commun.. 275. 582-588 (2000)

Hiroshi Nagai、Kyoko Takuwa、Masahiro Nakao、Emiko Ito、Masami Miyake、Masatoshi Noda、Terumi Nakajima：“箱形水母（海黄蜂）Carybdea rastoni 中的新型蛋白质毒素”Biochem。