Biochemical analysis of the enzymatic complex responsible for the generation of superoxide anion on the cell surface of Chattonella marina

对查托氏菌码头细胞表面产生超氧阴离子的酶复合物进行生化分析

• 批准号: 14560154
• 负责人: ODA Tatsuya
• 金额: $ 2.24万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560154/
• 关键词: Red Tide; Marine phytoplankton; Reactive Oxygen Species; Toxicity; Chattonella; Raphidophyceae; gycocalyx; NADPH oxidase; グリコキャッリックス

Chattonella. marina (C. marina), a raphidophycean flagellate, is one of the most noxious, red tide phytoplankton, and is highly toxic to fish, especially to yellowtail Seriola quinquer adiata. Previous our studies have demonstrated that a decrease in.oxygen partial pressure of arterial blood is the earliest physiological disturbance observed in fish after exposure to C. marina. Physiological and histological studies of fish exposed to C. marina also suggested that the blockade of respiratory water flow through the gill lamellae caused by excessive mucus interferes with oxygen transfer, resulting in asphyxia. One of the biochemical feature of C, marina is the ability to generate reactive oxygen species (ROS) such as superoxide anion and hydrogen peroxide. In this study, we analyzed the enzymatic complex. responsible for the production of ROS, which may be located on the cell-surface of C. marina. Our results suggest that glycocalyx may include ROS generating enzymatic system. Glycocalyx is a complex structure consisted of carbohydrates and proteins, and is easily discharged from C. marin cells in responding to various physical stimuli. In fact, our immunohistochemical analysis using anti-serum against glycocalyx revealed that significant amounts of glycocalyx were attached on the gill surface of yellowtail after C. marina exposure. Furthermore, we found that crude glycocalyx prepared from C. marina cell suspension produces ROS in the presence of NADPH. These findings suggest that glycocalyx may play a important role in the ROS-mediated fish-killing mechanism of C. marina. In addit ion to ROS, our recent study suggest that C. marina contains photoactive hemolytic toxin as a novel toxic factor. Further studies are required to clarify the involvement of such he molytic toxin in the fish-killing mechanism of C. marina.

查顿内拉marina（C. marina）是一种针形藻类鞭毛虫，是最具毒性的赤潮浮游植物之一，对鱼类，特别是黄尾五鳍鱼（Seriola quinquer adiata）具有很高的毒性。我们以前的研究已经证明，动脉血氧分压的降低是鱼在暴露于C后观察到的最早的生理紊乱。码头鱼类暴露于C.马里纳还指出，过多的粘液会阻碍呼吸水流通过鳃板，从而干扰氧气的输送，导致窒息。海洋梭菌的生化特性之一是产生活性氧（ROS），如超氧阴离子和过氧化氢。在这项研究中，我们分析了酶复合物。ROS的产生可能与C.码头我们的研究结果表明，糖萼可能包括活性氧产生酶系统。糖萼是一种由碳水化合物和蛋白质组成的复杂结构，很容易从C. marin细胞对各种物理刺激的反应。事实上，我们使用抗糖萼抗血清的免疫组织化学分析显示，在C.码头暴露。此外，我们还发现从C. marina细胞悬浮液在NADPH存在下产生ROS。这些结果表明，糖萼可能在C.码头我们最近的研究表明，C. marina含有光敏溶血毒素作为一种新的毒性因子。该毒素在C.码头

项目成果

Tamura, N.Tsuruta, K.Hirano, K.Yamaguchi, T.Oda: "Butyric acid sensitizes Vero cells to ricin-induced apoptosis via accelerated activation of multiple signal transduction pathways."Cell Struc.Func.. 28. 475-485 (2003)

Tamura、N.Tsuruta、K.Hirano、K.Yamaguchi、T.Oda：“丁酸通过加速激活多种信号转导途径，使 Vero 细胞对蓖麻毒素诱导的细胞凋亡敏感。”Cell Struc.Func.. 28. 475-485

S.Higuchi, T.Tamura, T.Oda: "Cross-talk between the pathways leading to the induction of apoptosis and the secretion of tumor necrosis factor-a in ricin-treated RAW 264.7 cells"J.Biochem.. 134. 927-933 (2003)

S.Higuchi、T.Tamura、T.Oda：“蓖麻毒素处理的 RAW 264.7 细胞中导致细胞凋亡诱导和肿瘤坏死因子 a 分泌的途径之间的串扰”J.Biochem.. 134. 927

Y.Iwamoto, H.Hideki, T.Oda, T.Muramatsu: "A study of tryptophan fluorescence quenching of bifunctional alginate lyase from a marine bacterium Pseudoalteromonas sp. strain No. 272 by acrylamide"Biosci.Biochechnol.Biochem.. 67. 1990-1992 (2003)

Y.Iwamoto、H.Hideki、T.Oda、T.Muramatsu：“丙烯酰胺对来自海洋细菌假交替单胞菌菌株 272 号的双功能藻酸盐裂解酶的色氨酸荧光猝灭的研究”Biosci.Biochechnol.Biochem.. 67。

Y.Sato, T.Oda, T.Muramatsu, Y.Matsuyama, T.Honjo: "Photosensitizing hemolytic toxin in Heterocapsa circularisquama, a newly identified harmful red tide dinoflagellate"Aquatic Toxicology. 56. 191-196 (2002)

Y.Sato、T.Oda、T.Muramatsu、Y.Matsuyama、T.Honjo：“圆形异壳藻中的光敏溶血毒素，一种新发现的有害赤潮甲藻”水生毒理学。

Y.Iwamoto, K.Iriyama, K.Osatomi, T.Oda, T.Muramatsu: "Primarystructure and chemical modification of some amino acid residues of bifunctional alginate lyase from a marine bacterium Pseudoalteromonas s strain No. 272."J.Protein Chem.. 21. 455-463 (2003)

Y.Iwamoto、K.Iriyama、K.Osatomi、T.Oda、T.Muramatsu：“来自海洋细菌假交替单胞菌 272 号菌株的双功能藻酸盐裂合酶的一些氨基酸残基的一级结构和化学修饰。”J.Protein Chem