Stopped-flow spectrometer for analysis of reaction kinetics

用于分析反应动力学的停流光谱仪

• 批准号: 10796272
• 负责人: Jeffrey S McFarlane
• 金额: $ 9.45万
• 依托单位: FORT LEWIS COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10796272
• 关键词: Anabolism; Bacteroides; Carboxy-Lyases; Clostridium; Coenzymes; Colon Carcinoma; Disease; Drug Design; Drug Targeting; Enzymes; Goals; Human; Kinetics; Malignant Neoplasms; Malignant neoplasm of pancreas; Metabolic Pathway; Oxidoreductase; Pathway interactions; Patients; Polyamines; Production; Public Health; Reaction; Spectrometry; Spermidine; Structural Models; Structure; Work; cancer cell; colon cancer progression; enzyme mechanism; gut bacteria; gut microbes; gut microbiome; kinetic model; new therapeutic target; therapeutic target

PROJECT SUMMARY Polyamine biosynthesis in human cancer cells is a long-standing therapeutic target. Accumulating evidence suggests that polyamine production by gut microbes modulates the progression of colon and pancreatic cancer. Biosynthesis of the polyamine spermidine by gut bacteria follows a pathway distinct from the human pathway making this bacterial pathway a unique drug target. The two central enzymes in this pathway, carboxyspermidine dehydrogenase and carboxyspermidine decarboxylase, have received only limited characterization. We propose to solve the first structure of a carboxyspermidine dehydrogenase and to develop kinetic and structural models describing the function of both carboxyspermidine dehydrogenase and carboxyspermidine decarboxylase in major gut constituents from the Bacteroides and Clostridium genera. Both carboxyspermidine dehydrogenase and decarboxylase use coenzymes with unique spectroscopic “handles” that can be interrogated by stopped- flow spectrometry to discern the kinetic steps of their respective catalytic cycles. A thorough structural and functional understanding of enzyme mechanisms is a necessary precursor to mechanism-guided drug design. This work will expand our fundamental understanding of a key gut microbe metabolic pathway: polyamine biosynthesis.

项目摘要 人类癌细胞中的多胺生物合成是长期的治疗靶点。积累证据 表明肠道微生物产生的多胺产生会调节结肠和胰腺癌的进展。 肠道细菌对多胺精子的生物合成遵循与人类途径不同的途径 使该细菌途径成为独特的药物靶标。该途径中的两个中心酶，羧培塞诗米丁 脱氢酶和羧培养地脱羧酶仅获得有限的特征。我们建议 解决羧培训酶脱氢酶的第一个结构，并开发动力学和结构模型 描述羧甲米定脱氢酶和羧甲米定脱羧酶的功能 主要的肠道构成杆菌和梭状芽胞杆菌属。两种羧氨基丁烷脱氢酶 脱羧酶使用辅酶具有独特的光谱“手柄”，可以通过停止来审问 - 流量光谱法辨别其各自催化循环的动力学步骤。彻底的结构和 对酶机制的功能理解是机理引导的药物设计的必要先驱。 这项工作将扩大我们对关键肠道微生物代谢途径的基本理解：多胺 生物合成。