X-ray Crystallographic Analysis of Diguanylate Cyclase Enzyme-Inhibitor Complexes

二鸟苷酸环化酶抑制剂复合物的 X 射线晶体分析

基本信息

批准号：
8582834
负责人：
Matthew B Neiditch
金额：
$ 0.53万
依托单位：
UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8582834
关键词：
Active Sites Address Affect Anti-Bacterial Agents Antibiotics Bacteria Bacterial Infections Binding Biochemical Cells Cessation of life Chemistry Clinical Collaborations Communities Complex Development Diffusion Enzyme Inhibitor Drugs Enzyme Inhibitors Eukaryota Exhibits Foundations Frequencies Future Genetic Genetic Screening Goals Human Immune response Immune system In Vitro Infection Intervention Laboratories Length Malignant Neoplasms Mediating Michigan Microbial Biofilms National Institute of Allergy and Infectious Disease Nutrient Organism Penetration Pharmaceutical Preparations Polymers Pseudomonas aeruginosa Research Research Design Roentgen Rays Role Salmonella typhimurium Second Messenger Systems Signal Pathway Signal Transduction Solutions Stress Structure Structure-Activity Relationship Surface System Thermodynamics Time Toxic effect United States National Institutes of Health Universities Vibrio Vibrio cholerae Water Work Yersinia pestis antimicrobial base bis(3&apos ,5&apos )-cyclic diguanylic acid conventional therapy diguanylate cyclase extracellular functional group genome analysis in vivo inhibitor/antagonist innovation killings pathogen phosphoric diester hydrolase public health relevance research study second messenger small molecule wasting

项目摘要

DESCRIPTION (provided by applicant): The long-term goals of this research are to explore the role of c-di-GMP in regulating bacterial signaling and to develop small molecules that interfere with c-di-GMP signaling pathways. C-di-GMP is a bacterial second messenger that commonly regulates biofilm formation in numerous human pathogens, including, among others, Vibrio sp., Salmonella typhimurium, Yersinia pestis, and Pseudomonas aeruginosa. Cellular levels of c-di-GMP are controlled by the opposing activities of diguanylate cyclases that synthesize c-di-GMP and phosphodiesterases that hydrolyze it. C-di-GMP, diguanylate cyclases, and c-di-GMP phosphodiesterases are not found in humans, making c-di-GMP signaling systems an attractive target for anti-bacterial intervention. Here we propose to determine X-ray crystal structures of diguanylate cyclases in complex with different digualylate cyclase inhibitors. The inhibitors were identified using a high-throughput in vivo genetic screen, demonstrated to specifically inhibit the enzymatic activity of multiple digualylate cyclases from different bacterial species in vitro, and shown to repress biofilm formation under static or flow conditions. The X-ray crystal structures will reveal the mechanistic basis of inhibitor function, e.g., whether the inhibitors are binding to the diguanylate cyclase active site and acting as competitive inhibitors, and also identify the inhibitor functional groups mediating the compound-target interactions. This information will drive structure-activity relationship studies designed t generate tighter binding antagonists through iterative rounds of compound redesign, synthesis, and in vivo genetic and in vitro biochemical analysis.

描述（由申请人提供）：这项研究的长期目标是探索C-DI-GMP在调节细菌信号传导中的作用，并开发出干扰C-DI-GMP信号传导途径的小分子。 C-DI-GMP是一种细菌的第二信使，通常调节许多人类病原体中的生物膜形成，包括颤音，鼠伤寒沙门氏菌，耶尔森氏菌，耶尔森氏菌和pseudomonas aeruginosa。 C-DI-GMP的细胞水平由合成C-DI-GMP和磷酸二酯酶的二甘氨酸酸环化酶的相反活性控制。 C-DI-GMP，二烷基酸酯循环酶和C-DI-GMP磷酸二酯酶在人类中找不到，使C-DI-GMP信号系统成为抗细菌干预的有吸引力的目标。在这里，我们建议确定与不同的digualylate旋转酶抑制剂复合物中二烷基酸环化酶的X射线晶体结构。使用体内遗传筛查的高通量鉴定抑制剂，该抑制剂在体外抑制了来自不同细菌种类的多种digualylate cyclase的酶活性，并在静态或流动条件下抑制生物膜形成。 X射线晶体结构将揭示抑制剂功能的机械基础，例如，抑制剂是否与二烷基酸环化酶活性位点结合并充当竞争性抑制剂，并识别介导复合目标相互作用的抑制剂官能团。该信息将驱动结构活性关系研究T设计t通过复合重新设计，合成以及体内遗传和体外生化分析的迭代回合产生更紧密的结合拮抗剂。