Disulfide redox balance in Schistosoma mansoni

曼氏血吸虫的二硫键氧化还原平衡

• 批准号: 6596463
• 负责人: DAVID LEE WILLIAMS
• 金额: $ 14万
• 依托单位: ILLINOIS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-02 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6596463
• 关键词: NAD(P)H oxidoreductase; SDS polyacrylamide gel electrophoresis; Schistosoma mansoni; antioxidants; drug design /synthesis /production; enzyme activity; high performance liquid chromatography; host organism interaction; ion exchange chromatography; laboratory mouse; liquid chromatography mass spectrometry; oxidative stress; peptide chemical synthesis; polymerase chain reaction; protein purification; recombinant proteins; schistosomiasis; sulfides; thioredoxin; western blottings; NAD(P)H oxidoreductase; SDS polyacrylamide gel electrophoresis; Schistosoma mansoni; antioxidants; drug design /synthesis /production; enzyme activity; high performance liquid chromatography; host organism interaction; ion exchange chromatography; laboratory mouse; liquid chromatography mass spectrometry; oxidative stress; peptide chemical synthesis; polymerase chain reaction; protein purification; recombinant proteins; schistosomiasis; sulfides; thioredoxin; western blottings

DESCRIPTION (provided by the applicant): Schistosomiasis is an important tropical parasitic human and veterinary disease. Although an inexpensive and highly effective anti-schistosome drug is in use 20 million individuals still suffer severe disease symptoms, transmission rates have changed little and there is evidence for the development of drug resistant parasites. Because there is currently no suitable alternative therapy available, there is an urgent need for the development of novel chemotherapeutic agents. Schistosomes in their definitive and intermediate hosts must be able to survive in the presence of immune and self generated reactive oxygen compounds and provide disulfide-reducing equivalents for a number of critical enzymatic pathways. Two parallel detoxification/disulfide reduction systems occur in most organisms, one based on glutathione and the other based on thioredoxin. Recent results indicate that adult Schistosoma mansoni are deficient in glutathione reductase and thioredoxin reductase key enzymes in these pathways. Instead these activities are found together in a single protein thioredoxin glutathione reductase. This application proposes to investigate the role of thioredoxin glutathione reductase in parasite antioxidant defense and redox balance. The native protein will be purified and biochemically characterized. Active recombinant protein will be produced in order to carry out more extensive analysis. The role of the protein in redox defenses in vitro and in vivo will be investigated using known enzyme inhibitors. The stage and tissue expression of the enzyme will be determined. Because disulfide redox balance in schistosomes is centered on a single, key enzyme and is fundamentally different from host mechanisms we propose that this pathway is a promising target for novel, rational drug design.

描述（由申请人提供）：血吸虫病是一种重要的热带寄生虫和兽医疾病。尽管使用廉价且高效的抗速剂药物正在使用2000万个人仍然患有严重的疾病症状，但传播率几乎没有变化，并且有证据表明耐药性寄生虫的发展。由于目前尚无合适的替代疗法，因此迫切需要开发新型的化学治疗剂。其确定性和中间宿主中的血吸虫必须能够在存在免疫和自我产生的活性氧化合物的情况下生存，并为许多关键的酶促途径提供二硫键的等效物。大多数生物体发生了两个平行的排毒/二硫化物还原系统，一种基于谷胱甘肽，另一个基于硫氧还蛋白。最近的结果表明，在这些途径中，成年曼氏菌缺乏缺乏谷胱甘肽还原酶和硫氧还蛋白还原酶键酶。相反，这些活性在单个蛋白硫氧还蛋白谷胱甘肽还原酶中一起发现。该应用建议研究硫氧还蛋白谷胱甘肽还原酶在寄生虫抗氧化剂防御和氧化还原平衡中的作用。天然蛋白将被纯化并在生化表征。将产生活跃的重组蛋白，以进行更广泛的分析。该蛋白在体外和体内的氧化还原防御性中的作用将使用已知的酶抑制剂研究。将确定酶的阶段和组织表达。由于血块中的二硫键氧化还原平衡集中在一个关键酶上，并且与宿主机制根本不同，因此我们建议这种途径是新颖的，理性药物设计的有希望的目标。