MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF A SECRETED LIPASE IN LEISHMANIA

利什曼原虫分泌的脂肪酶的分子和生物化学特征

• 批准号: 7381355
• 负责人: ALISON M SHAKARIAN
• 金额: $ 10.44万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381355
• 关键词: MOLECULAR; BIOCHEMICAL; CHARACTERIZATION; SECRETED; LIPASE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The proposed research is designed to contribute to the understanding of mechanisms of Leishmania survival, growth and development. The disease spectrum of leishmaniasis ranges from mild self-limiting cutaneous ulcers to a potentially fatal visceral infection, each dependant on the species of infectious parasite. Specific proteins secreted by Leishmania enable the organisms to sense, respond to, and alter their environment. Such secreted proteins are thus potential targets for the development of new compounds to prevent or treat this diseases. Currently there is no vaccine available for the prevention of transmission of these parasites. Toxic antimony compounds have been used unsuccessfully. Lipases are lipolytic enzymes that hydrolyze the ester bond of triglycerides. In other systems, lipase activity is involved in the reorganization of membrane structure, cell signaling or nutrient acquisition. These observations suggest that lipase activity is essential for Leishmania although presently the biological significance of this enzyme has not been proven experimentally. The working hypothesis, that lipase is essential to Leishmania, will be tested. The following approaches to determine the role of this enzyme in the parasite life cycle are planned: 1) The full-length copy of the lipase gene will be identified and characterized; the expression of lipase will be examined though out the parasite developmental life cycle and the phylogenic conservation of lipase among kinetoplastid protozoa will be determined. 2) Null-mutants deficient in lipase activity will be generated and tested for viability. 3) The lipase gene will be expressed as a recombinant protein and assayed for substrate preference. In addition, various classes of organophosphate inhibitors of lipase will be tested to determine the sensitivity of the leishmanial lipase to these compounds.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。拟议的研究旨在有助于理解利什曼尼亚生存，生长和发展的机制。利什曼病的疾病范围从轻度的自限制皮肤溃疡到潜在的致命内脏感染，每种溃疡都取决于感染性寄生虫的种类。利什曼原虫分泌的特定蛋白质使生物体能够感知，反应和改变其环境。因此，这种分泌的蛋白质是开发新化合物以预防或治疗这种疾病的潜在靶标。目前尚无疫苗可预防这些寄生虫的传播。有毒仇敌化合物未成功使用。脂肪酶是水解甘油三酸酯的酯键的脂肪酶。在其他系统中，脂肪酶活性参与膜结构，细胞信号传导或养分的重组。这些观察结果表明，脂肪酶活性对于利什曼原虫至关重要，尽管目前尚未通过实验证明该酶的生物学意义。脂肪酶对利什曼原虫至关重要的工作假设将进行测试。计划了以下方法来确定该酶在寄生虫生命周期中的作用：1）将确定和表征脂肪酶基因的全长副本；尽管将确定寄生虫发育生命周期，并且将确定脂肪酶的表达，并确定脂肪酶对动力学原生动物中脂肪酶的系统发作。 2）将生成并测试缺乏脂肪酶活性的无效脂肪。 3）脂肪酶基因将表示为重组蛋白，并分析底物偏好。此外，将测试各种脂肪酶的有机磷酸盐抑制剂，以确定利什曼脂肪酶对这些化合物的敏感性。