Glucose Sensing and Hexokinases in the African Trypanosome

非洲锥虫中的葡萄糖传感和己糖激酶

• 批准号: 9261578
• 负责人: JAMES Culvin MORRIS
• 金额: $ 24.31万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9261578
• 关键词: Africa South of the Sahara; African; African Trypanosomiasis; Area; Biochemical; Biology; Blood Circulation; Cell physiology; Cellular biology; Centers of Research Excellence; Collaborations; Complex; Core Facility; Cues; Data; Detergents; Development; Disease; Distant; Ectopic Expression; Environment; Enzymes; Funding; Generations; Genetic; Glucose; Glycolysis; Glycosome; Goals; Hexokinase 2; Human; Image; Immunochemistry; Infection; Insecta; Laboratories; Life Cycle Stages; Light; Mammals; Measures; Mediating; Mentors; Mentorship; Metabolism; Microscopy; Mission; Mitochondria; Molecular; Nutrient; Organelles; Outcome; Parasites; Pathway interactions; Process; Proteins; Public Health; Publications; Regulation; Research; Research Infrastructure; Research Personnel; Resolution; Risk; Role; Testing; Trypanosoma; Trypanosoma brucei brucei; Tsetse Flies; United States National Institutes of Health; Universities; Voltage-Dependent Anion Channel; Work; base; enzyme activity; glucose metabolism; hexokinase; innovation; interest; kinetosome; matriculation; neglected tropical diseases; new therapeutic target; pathogen; peroxisome; programs; response; therapeutic development; unpublished works

PROJECT SUMMARY/ABSTRACT The infectious lifecycle stage of the African trypanosome, Trypanosoma brucei, relies exclusively on glycolysis for ATP generation. The parasite responds to dynamic environmental cues to regulate this pathway. The goal of this application is to resolve the mechanisms that regulate localization of the essential glycolytic enzymes, T. brucei hexokinase 1 and 2, with a particular emphasis on understanding the glucose-sensitive subcellular localization of the proteins. We have previously demonstrated that T. brucei hexokinase 2, which was heretofore believed to be limited in its distribution to a peroxisome-like organelle called the glycosome, has in addition a life-cycle dependent extra-glycosomal localization. In the bloodstream form parasites, the protein was found in the flagellum while in the insect stage the protein was found proximal to the basal bodies. Our preliminary data suggests that the localization is dependent on environmental glucose availability and that the hexokinases are associated with a detergent-insoluble fraction of the mitochondria. Using both ectopic expression of tagged proteins and immunochemistry in collaboration with microscopy and biochemical approaches, we will identify the enzyme domains that are required for localization. Additionally, we will begin to resolve the mechanisms required for glucose-dependent localization, with a particular focus on the association of the hexokinases with the parasite mitochondrion. Through these studies, new ways of targeting glucose metabolism, an essential parasite pathway, will be identified.

项目概要/摘要 非洲锥虫布氏锥虫的感染生命周期阶段完全依赖于糖酵解 用于 ATP 生成。寄生虫响应动态环境线索来调节该途径。目标 该应用的目的是解决调节必需糖酵解酶 T 定位的机制。 brucei 己糖激酶 1 和 2，特别强调了解葡萄糖敏感的亚细胞 蛋白质的定位。我们之前已经证明了 T. brucei 己糖激酶 2 迄今为止，人们认为其分布仅限于称为糖体的过氧化物酶体样细胞器， 另外还有生命周期依赖性的糖体外定位。在血液中形成寄生虫，蛋白质 在鞭毛中发现，而在昆虫阶段，该蛋白质被发现靠近基体。我们的 初步数据表明，定位取决于环境葡萄糖的可用性，并且 己糖激酶与线粒体的去污剂不溶部分相关。使用两种异位 与显微镜和生物化学合作进行标记蛋白和免疫化学的表达 方法，我们将确定定位所需的酶结构域。此外，我们将开始 解决葡萄糖依赖性定位所需的机制，特别关注关联 己糖激酶与寄生虫线粒体的结合。通过这些研究，靶向葡萄糖的新方法 将确定新陈代谢这一重要的寄生虫途径。