Study on 14-3-3 related proteins and signal transduction for control of African tyrpanosomiasis.

控制非洲锥虫病的14-3-3相关蛋白及信号转导研究。

• 批准号: 14570228
• 负责人: FUKUMA Toshihide
• 金额: $ 2.24万
• 依托单位: KURUME 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-14570228/
• 关键词: 14-3-3; Trypanosoma b.brucei; dsRNAi; pQuadra; African trypanosomiasis; therapeutic target; therapeuticc target; Trypanosoma brucei

Protein 14-3-3 was described to be rich in CNS and ubiquitously found in eukaryotic organisms of plants and animals. Quite a many proteins have been reported to bind to 14-3-3 then it is estimated the protein has important roles in the cell cycle, apoptosis and other signal transduction paths.We examined 14-3-3 of Trypanosoma b.brucei (Tb) and remark the followings.1) The genes of 2 isotypes (I, II) of 14-3-3 could be cloned. The result produced the recombinant proteins and sensitive mono-specific rat antibodies against them.2) Though 14-3-3 distinctive motifs are conserved, the characteristics of trypanosome 14-3-3 are quite different form those of mammals.3) We developed solely effective vector construct named pQuadra (pQ) to perform dsRNAi without leakage before induction.4) Procyclic forms (pr) were examined by dsRNAi on 14-3-3 applying pQ. Morphologically pr was damaged to bizarre multiflagellated form and disoriented kinetoplsts or nuclei delivery. The growth of pr was suppressed.5)On blood stream forms (bs) after induction of dsRNAi with pQ more drastic effect appeared earlier than pr and the growth was entirely suppressed6) The morphological change of bs was very similar to the effect of okadaic acid addition to culture of bs, which is protein phosphates (pp) inhibitor. Analysis based on relation between 14-3-3 and pp we came very close to the therapeutic target of African trypanosomiasis.

蛋白质 14-3-3 富含中枢神经系统，普遍存在于植物和动物的真核生物中。已有相当多的蛋白质与14-3-3结合，推测该蛋白质在细胞周期、细胞凋亡和其他信号转导途径中具有重要作用。我们检测了Trypanosoma b.brucei (Tb)的14-3-3并进行了如下评论。1)可以克隆14-3-3的2个同种型(I、II)的基因。结果产生了重组蛋白和针对它们的敏感单特异性大鼠抗体。2) 尽管 14-3-3 独特基序是保守的，但锥虫 14-3-3 的特征与哺乳动物的特征有很大不同。3) 我们开发了唯一有效的载体构建体，名为 pQuadra (pQ)，用于在诱导前进行 dsRNAi，而不会泄漏。4) 通过以下方法检查了原环形式 (pr)： 应用 pQ 对 14-3-3 进行 dsRNAi。形态学上，pr 被破坏为奇怪的多鞭状形式，动质体或细胞核输送失去方向性。 pr的生长受到抑制。5)用pQ诱导dsRNAi后，对血流形式(bs)的影响比pr更早出现，并且生长完全受到抑制6)bs的形态变化与向bs培养物中添加冈田酸的效果非常相似，冈田酸是蛋白磷酸盐(pp)抑制剂。基于14-3-3和pp之间关系的分析，我们非常接近非洲锥虫病的治疗靶点。

项目成果

福間利英: "病原性自由生活アメーバ感染による髄膜脳炎の症例・診断・治療〜PCRの有用性〜"科学療法の領域. 19. 51-58 (2003)

Toshihide Fukuma：“致病性自由生活阿米巴感染引起的脑膜脑炎的病例、诊断和治疗〜PCR的用途〜”科学治疗领域19. 51-58（2003）。

R.E.Morales, et al.: "Infection and dissemination of two dengue type-2 viruses isolated from patients exhibiting different disease severity in orally infected Aedes aegypti form different geographic origin."Med.Entomol.Zool.. 53. 21-27 (2002)

R.E.Morales 等人：“从不同地理来源的口腔感染埃及伊蚊中表现出不同疾病严重程度的患者分离出的两种登革热 2 型病毒的感染和传播。”Med.Entomol.Zool.. 53. 21-27 (2002

R.E.Morales, et al.: "Infection and dissemination of two dengue type-2 viruses isolated from patients exhibiting different disease severity in orally infected Aedes aegypti form different geographic origin."Med.Entomol. Zool.. 53. 21-27 (2002)

R.E.Morales 等人：“从不同地理来源的口腔感染埃及伊蚊中表现出不同疾病严重程度的患者分离出两种登革热 2 型病毒的感染和传播。”Med.Entomol。

E.Nyarko, et al.: "Toxic effects of mercury(II), cadmium(II)and lead(II) porphyrins on Trypanosoma brucei brucei growth."Chemico-Biological Interacions. 139. 177-185 (2002)

E.Nyarko 等人：“汞 (II)、镉 (II) 和铅 (II) 卟啉对布氏锥虫生长的毒性作用。”化学-生物相互作用。

Elvis Nyarko, et al.: "In vitro toxicity of palladium(II) and gold(III) porphyrins and their aqueous metal ion counterparts on Trypanosoma brucei brucei growth."Chemico-Biological Interacions. (in press). (2004)

Elvis Nyarko 等人：“钯 (II) 和金 (III) 卟啉及其水溶液金属离子对应物对布氏锥虫生长的体外毒性。”化学-生物相互作用。