Trehalose-6-phosphate phosphatase: a target for anti-onchocerciasis therapeutics

海藻糖-6-磷酸磷酸酶：抗盘尾丝虫病治疗的靶点

• 批准号: 8606399
• 负责人: Karen N. Allen
• 金额: $ 19.68万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-18 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606399
• 关键词: Active Sites; Affect; Africa; Americas; Anabolism; Area; Asia; Binding; Binding Sites; Caenorhabditis elegans; Catalytic Domain; Cessation of life; Complement; Country; Crystallization; Culicidae; Development; Disease; Drug Targeting; Electrostatics; Encapsulated; Endemic Diseases; Energy-Generating Resources; Enzymes; Escherichia coli; Family; Filarial Elephantiases; Foundations; Funding; Genes; Genome; Goals; Grant; Homologous Gene; Human; Individual; Infection; Intervention; Knowledge; Laboratories; Lead; Life Cycle Stages; Ligand Binding; Ligands; Ligation; Link; Low Income Population; Lymphatic; Measures; Modeling; Molecular Conformation; Nematoda; Onchocerciasis; Organism; Orthologous Gene; Parasitic nematode; Pathway interactions; Pharmaceutical Preparations; Phenotype; Phosphoric Monoester Hydrolases; Population; Probability; Recombinant Proteins; Recombinants; Research; Research Personnel; Resolution; Roentgen Rays; Scheme; Severities; Site; Stress; Structure; Structure-Activity Relationship; Sucrose; Surface; Therapeutic; Therapeutic Intervention; Toxic effect; Trehalose; Vanadates; Work; aluminum fluoride; analog; base; beryllium fluoride; conformer; design; drug development; drug discovery; enzyme structure; experience; inhibitor/antagonist; inorganic phosphate; man; member; mutant; protein structure; public health relevance; research study

DESCRIPTION (provided by applicant): Parasitic nematodes are responsible for numerous chronically incapacitating and deforming diseases in Africa, Asia, and the Americas. Among these diseases is lymphatic filariasis, which is a mosquito-transmitted disease, endemic to 81 countries. It is estimated that 120 million people are infected with this disease. Enzymes that are essential for the parasitic nematodes but that do not have a human homologue are potential drug targets for therapeutic intervention. The availability of the genome from B. malayi, the representative organism for filarial nematodes, has enabled the ranking of potential drug targets from this parasitic organism. One such enzyme is trehalose-6-phosphate phosphatase (T6PP), which is required for the biosynthesis of trehalose. The oblation of T6PP activity in the C. elegans model commonly used for parasitic nematodes ultimately leads to organism death, probably due to the accumulation of trehalose 6-phosphate (T6P). Because T6PP is a member of the haloalkanoate dehalogenase superfamily of phosphatases, knowledge about the structure/function relationships in this family can be used to define T6PP for drug development. The objective of the proposed study is to identify the steric and electrostatic features of the T6PP active-site region that can be exploited in the design of lead inhibitors. Because the two domains of the enzyme can rotate to open the active site for ligand exchange, the surface area that can be potentially targeted by an inhibitor will be obtained by determining structures of bot open and closed conformers. The research plan is focused on a single Aim: Define the Target Site for the Development of Drug-like Inhibitors of B. malayi Trehalose-6-phosphate Phosphatase. The ¿lead¿ T6PP X-ray crystal structure will be determined using the recombinant enzyme from B. malayi and C. elegans to define the overall structure of the protein. Co-crystallization and crystal soaking experiments will capture the closed state (using an inactive T6PP mutant and T6P or using T6PP plus an inert substrate analog) and transition-state conformations (T6PP and trehalose plus vanadate, beryllium fluoride, or aluminum fluoride). Analysis of structure-activity relationships will be used to define the binding interactions which dominate the contributions to the ligand binding energy. The structure of the enzyme in the cap-open conformation will be determined (using apo T6PP and T6PP domain-domain binding mutants) in order to design and evaluate bidentate inhibitors that can fill and complement the expanded binding crevice of the cap-open conformer. This work will deliver the foundation for the development of a drug for the treatment of disease(s) inflicting the large segment of the world's population suffering from infection by parasitic nematodes.

描述（由适用提供）：寄生线虫负责在非洲，亚洲和美洲的许多长期丧失能力和变形疾病。这些疾病中有淋巴丝虫病，这是一种蚊子传播的疾病，对81个国家。据估计，有1.2亿人感染了这种疾病。对寄生线虫至关重要的酶，但没有人类同源物是治疗干预的潜在药物靶标。来自丝状线虫的代表生物马来语的基因组的可用性使该寄生生物的潜在药物靶标有能力排名。一种这样的酶是海藻糖6-磷酸磷酸酶（T6PP），这是天虫的生物合成所必需的。通常用于寄生线虫的C.秀丽隐杆线虫模型中T6PP活性的目的最终导致生物体死亡，这可能是由于三磷酸三磷酸盐（T6P）的积累。由于T6pp是磷酸酶的卤代甲酯脱核酶超家族的成员，因此可以使用该家族中的结构/功能关系来定义T6pp进行药物开发。拟议的研究的目的是确定可以在铅抑制剂设计中探索的T6PP活性位点区域的空间和静电特征。由于酶的两个结构域可以旋转以打开配体交换的活性位点，因此可以通过确定机器人开放和封闭构象体的结构来获得可能由抑制剂靶向的表面积。该研究计划的重点是一个目标：定义靶位点，用于开发马来链球菌-6-磷酸二磷酸杆菌的药物样抑制剂。使用来自马来芽孢杆菌和秀丽隐杆线虫的重组酶来确定铅XPP X射线晶体结构，以定义蛋白质的整体结构。共结晶和晶体浸泡实验将捕获封闭状态（使用无活跃的T6PP突变体和T6P，或使用T6PP加上惰性底物类似物）和过渡状态构象（T6PP和T6PP和T6PP和T6PP，以及脱甲酸盐，孢子虫，孢子菌，氟化氟化物或铝氟化物）。结构活性关系的分析将用于定义主导对配体结合能的贡献的结合相互作用。为了设计和评估可以填充和补充帽盖构象的扩展结合缝隙，将确定盖构构中酶的结构（使用APO T6PP和T6PP结构域结合突变体）。这项工作将为开发一种药物的基础，以治疗疾病，从而造成寄生虫感染的世界大部分人口。