Biologically Active Key Compounds of Protozoan Ciliates

原生动物纤毛虫的生物活性关键化合物

• 批准号: 12045256
• 负责人: IIO Hideo
• 金额: $ 8.19万
• 依托单位: Osaka City University
• 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-12045256/
• 关键词: ciliate; Blepharisma; conjugation; defense toxin; Climacostomum; climacostol; Spirostomum; spirostomin; スピロストマム; スピロストモン; ガモン; 糖タンパク質; アスパラギン結合型糖鎖; ブレファルモン

1. Conjugation of ciliate Blepharisma japonicum Conjugation in Blepharisma japonicum is induced byinteraction between complementary mating-types I and II, which excrete blepharmone (gamone 1) and blepharismone (gamone 2), respectively. Gamone 1 transforms type II cells such that they can unite, and gamone 2 similarly transforms type I cells. The partial amino acid sequence of four peptides obtained from gamone I was determined. By using mRNA from mating type, gamone 1 cDNA was isolated by rapid amplification. The cDNA sequence contains an ORF of 305 amino acids and codes a novel protein. We also estimated that the sugar sequence for the glycoprotein consisted from four N-acetylglucosamines and three mannoses.To elucidate the receptor of blepharismone, a photoaffinity label probe of blepharismone is designed. We synthesized a model compound Nbenzoyl blepharismone based on the strategy developed before. The model compbund did not have mating inducing activity as expected, it was found to … More have the significant activity ofmating inhibition.2-1. Defense toxins of Climacostomum Virens The heterotrich ciliate Climacostomum virens has colourless cortical granules morphologically similar to pignentocysts (extrusive pigment granules) in the two coloured heterotrichs, Blepharisma japonicum and Stentor coeruleus, which are known to function as defensive organelles against predators. Climacostol, a defense toxin of the heterotrich ciliate Climacostomum virens was isolated and the structure was established as 5-(Z)-non-2-enyl-benzene-1, 3-diol. The structure was rigorously confirmed by the total synthesis. The two congeners of climacostol contained in this ciliate were determined as 5-(Z, Z, Z)-undeca-2, 5, 8-trienyl-benzene-I, 3-diol and 5-(Z, Z)-undeca-2, 5-dienyl-benzene-1, 3-diol.2-2. Defense toxin of Spirostomum teres A heterotrich ciliate Spirostomum teres has colorless cortical granules that are mophologically similar to the pigment granules of B. japonicum and S. coeruleus, and the predatory ciliate D. margaritifer rarely attacks S. teres. We found the ethanol extracts of S. teres shown to be toxic to D. margaritifer. The structure of a toxic compound named spirostomin was elucidated as spiro-2,5 -dimethyl-5,6,7,8-tetrahydronaphtalene-1,4-dione-8,6'-pyran-2',5'-dione. Less

1.日本小眼纤毛虫的偶联作用日本小眼纤毛虫的偶联作用是由互补交配型Ⅰ和Ⅱ之间的相互作用诱导的，它们分别分泌小眼激素（配子体1）和小眼激素（配子体2）。配子1转化II型细胞，使它们能够结合，配子2类似地转化I型细胞。测定了从gamone I中获得的四种肽的部分氨基酸序列。以交配型的mRNA为模板，通过快速扩增法分离配子体1的cDNA。该cDNA序列含有305个氨基酸的ORF，编码一种新的蛋白质。为了阐明白藜芦醇的受体，我们设计了一种白藜芦醇的光亲和标记探针。在此基础上，我们合成了一个模型化合物N-苯甲酰基苯甲酮。模型化合物没有预期的交配诱导活性，发现其 ...更多信息 具有显著的交配抑制活性.绿变口线虫的防御毒素异毛类纤毛虫绿变口线虫具有无色的皮层颗粒，形态上类似于两种有色异毛类（Blepharisma japonicum和Stentor coeruleus）中的pignentocytes（喷出色素颗粒），已知其作为防御性细胞器对抗捕食者。从异毛类纤毛虫绿色爬口虫Climacostomum virens中分离得到一种防御毒素Climacostomium，其结构经鉴定为5-（Z）-壬-2-烯基苯-1，3-二醇。其结构通过全合成得到了严格的证实。经鉴定，该纤毛虫中含有2种异环戊烯类化合物，分别为5-（Z，Z，Z）-十一碳-2，5，8-三烯基苯-1，3-二醇和5-（Z，Z）-十一碳-2，5-二烯基苯-1，3-二醇。圆口线虫的防御毒素一种异毛类纤毛虫圆口线虫具有无色的皮层颗粒，其形态与B的色素颗粒相似。japonicum和S. coeruleus和捕食性纤毛虫D. margaritifer很少攻击S.圆肌我们发现S.圆线虫对D.玛格丽特。一种有毒化合物spirostomin的结构被鉴定为螺-2，5-二甲基-5，6，7，8-四氢萘-1，4-二酮-8，6 '-吡喃-2'，5 '-二酮。少

项目成果

Y.Muto, Y.Tanabe, K.Kawai, H.Iio: "Inhibition of mitochondrial respiration by climacostol"Jpn. J. Protozool.. 36(1). 25-26 (2003)

Y.Muto、Y.Tanabe、K.Kawai、H.Iio：“climacostol 对线粒体呼吸的抑制”Jpn。

Miyake,M.,Harumoto,T.,Iio,H.: "Defense function of pigment granules in Stentor coeruleus"Europ.J.Protistol.. (in press). (2001)

Miyake,M.,Harumoto,T.,Iio,H.：“Stentor coeruleus 色素颗粒的防御功能”Europ.J.Protistol..（出版中）。

A.Miyake, F Buonnano, P.Saltalamachia, M.E.Masaki, H.Iio: "Chemical defence by means of extrusive cortical granules in the heterotrich ciliate Climacostomum virens."Europ.J.Protistol.. 30. 25-36 (2003)

A.Miyake、F Buonnano、P.Saltalamachia、M.E.Masaki、H.Iio：“通过异毛纤毛虫 Climacostum virens 中的突出皮质颗粒进行化学防御。”Europ.J.Protistol.. 30. 25-36 (2003)

M.E.Masaki, S.Hiro, Y.Usuki T.Harumoto, M.N.Terazima, E.Buonnano, A.Miyake, H.Iio: "Climacostol, A Defense Toxin of Climacostomum virens (Protozou, Ciliata), and It's Congeners."Tetahedron. (in press).

M.E.Masaki、S.Hiro、Y.Usuki T.Harumoto、M.N.Terazima、E.Buonnano、A.Miyake、H.Iio：“Climacostol，Climacostum virens（Protozou，Ciliata）的防御毒素，及其同类物。”

M.E.Masaki, S.Hiro, Y.Usuki T Hanunoto, M.N.Terazima, F.Buonnano, A.Miyake, H.ho: "Climacostol, A Defense Toxin of Climacostomum virens (Protozoa, Ciliata), and It's Congeners."Tetrahedron. (In press).

M.E.Masaki、S.Hiro、Y.Usuki T Hanunoto、M.N.Terazima、F.Buonnano、A.Miyake、H.ho：“Climacostol，Climacostum virens（原生动物、纤毛虫）的防御毒素，及其同类物。”四面体。