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: 8427651
• 负责人: Karen N. Allen
• 金额: $ 25.38万
• 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 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8427651
• 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（由申请人提供）：寄生线虫导致非洲、亚洲和美洲的多种慢性致残和畸形疾病，其中淋巴丝虫病是一种蚊子传播的疾病，据估计在 81 个国家流行。 1.2 亿人感染了这种疾病，而寄生线虫所必需的但没有人类同源物的酶是潜在的药物靶标。丝虫线虫的代表性生物马来丝虫的基因组的可用性使得能够对这种寄生生物的潜在药物靶标进行排序，其中一种酶是海藻糖-6-磷酸磷酸酶（T6PP），这是必需的。用于海藻糖生物合成的线虫模型中 T6PP 活性的消除最终可能导致生物体死亡。由于海藻糖 6-磷酸 (T6P) 的积累，由于 T6PP 是磷酸酶卤代烷酸脱卤酶超家族的成员，因此有关该家族结构/功能关系的知识可用于定义 T6PP 的药物开发目标。拟议的研究旨在确定 T6PP 活性位点区域的空间和静电特征，这些特征可用于先导抑制剂的设计，因为酶的两个结构域可以。旋转以打开配体交换的活性位点，通过确定 bot 开放构象异构体和闭合构象异构体的结构，将获得抑制剂可能靶向的表面积。马来芽胞杆菌海藻糖-6-磷酸磷酸酶类药物抑制剂的开发。带领T6PP X 射线晶体结构将使用来自马来芽胞杆菌和秀丽隐杆线虫的重组酶来确定蛋白质的整体结构，共结晶和晶体浸泡实验将捕获闭合状态（使用非活性 T6PP 突变体和 T6P）。或使用 T6PP 加惰性底物类似物）和过渡态构象（T6PP 和海藻糖加钒酸盐、氟化铍或铝）结构-活性关系的分析将用于定义对配体结合能的主导贡献的结合相互作用。将确定酶在开帽构象中的结构（使用apo T6PP和T6PP结构域-结构域）。结合突变体），以设计和评估可以填充和补充开帽构象异构体扩大的结合间隙的双齿抑制剂。这项工作将为开发用于治疗造成疾病的药物奠定基础。世界上很大一部分人口遭受寄生线虫感染。