Vulnerabilities in Metabolite, Heme-lron and Redox Environments

代谢物、血红素铁和氧化还原环境中的漏洞

• 批准号: 8724066
• 负责人: Celia Goulding
• 金额: $ 6.07万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2013-09-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8724066
• 关键词: Address; Alanine; Amidohydrolases; Anabolism; Antibiotic Resistance; Antibiotics; Autolysis; Bacillus (bacterium); Binding; Biochemistry; Biogenesis; Carbapenems; Carboxypeptidase; Cell Wall; Cell division; Cells; Collaborations; Complex; Cues; Cycloserine; Drug Targeting; Environment; Enzymes; Ethambutol; Genetic; Genetic Structures; Genus Mycobacterium; Growth; Health; Heme; Homeostasis; Human Development; Hydrolase; Immunology; Instruction; Knowledge; Lactamase; Ligands; Lipids; Methods; Microbiology; Molecular; Molecular Conformation; Multienzyme Complexes; Mycobacterium tuberculosis; Oxidation-Reduction; Pathway interactions; Peptides; Production; Proteins; Regulation; Regulatory Pathway; Research Personnel; Role; Signal Pathway; Staging; Structure; Substrate Specificity; Sulfur; Testing; Time; Toxic effect; Work; amidase; analog; base; cell envelope; cell growth; chemical genetics; design; inhibitor/antagonist; innovation; isoniazid; killings; member; multidisciplinary; mycobacterial; novel; novel therapeutics; programs; protein protein interaction; small molecule; sugar; tuberculosis drugs

This project is focused on defining new mechanistic paradigms for Mycobacterium tuberculosis (Mtb) cell wall. biosynthesis and remodeling, which are essential for cell growth and division. Cell-wall biosynthesis is the target of well-known, critical anti-tuberculars, including isoniazid, cycloserine and ethambutol. We will concentrate on the pepfidoglycan (PG) layer of the cell wall, which serves as a meshwork for the structural integrity of the bacillus. Recent progress has identified proteins involved in PG homeostasis either enzymatically (PG hydrolases) or in regulatory roles (PknB, FhaA, and the lipid II flippase). These, along with other cell-wall biosynthetic enzymes, represent potential vulnerabilities that could be exploited for design of new TB drugs. We will take a multidisciplinary, multi-investigator approach enabled by our Core capabilities to address major questions about Mtb cell-wall biogenesis and its regulation. In Aim 1, we will define new molecular mechanisms of auto-inhibition of PG hydrolases to discover how their toxicity is mitigated. In Aim 2, we will structurally characterize complexes of active PG hydrolases. To uncover indirect cell-wall vulnerabilities, we also will determine the structures of complexes of regulatory factors that control PG biosynthesis in diverse environments. These studies will uncover for the first time activation mechanisms that control PG integrity. Aim 3 focuses on determining structures of small-molecule complexes of PG hydrolases and other cell-wall targets to define the basis for subrate- and inhibitor-binding specificty. By testtng fundamental hypotheses about cell-wall biosynthettc pathways and regulatory networks, this project sets the stage to develop potent, selective inhibitors of Mtb growth.

该项目的重点是定义结核分枝杆菌 (Mtb) 细胞壁的新机制范例。 生物合成和重塑，这对于细胞生长和分裂至关重要。细胞壁生物合成是众所周知的关键抗结核药物的目标，包括异烟肼、环丝氨酸和乙胺丁醇。我们将重点关注细胞壁的肽聚糖（PG）层，它作为芽孢杆菌结构完整性的网络。最近的进展已经确定了参与 PG 稳态的蛋白质，无论是酶促作用（PG 水解酶）还是调节作用（PknB、FhaA 和脂质 II 翻转酶）。这些与其他细胞壁生物合成酶一起代表了可用于设计新结核病药物的潜在漏洞。 我们将采用由我们的核心能力支持的多学科、多研究者方法来解决有关结核分枝杆菌细胞壁生物发生及其调控的重大问题。在目标 1 中，我们将定义 PG 水解酶自动抑制的新分子机制，以发现如何减轻其毒性。在目标 2 中，我们将对活性 PG 水解酶复合物进行结构表征。为了发现间接的细胞壁脆弱性，我们 还将确定在不同环境中控制 PG 生物合成的调节因子复合物的结构。这些研究将首次揭示控制 PG 完整性的激活机制。 目标 3 侧重于确定 PG 水解酶和其他细胞壁靶标的小分子复合物的结构，以确定亚速率和抑制剂结合特异性的基础。 通过测试有关细胞壁生物合成途径和调控网络的基本假设，该项目为开发有效的、选择性的结核分枝杆菌生长抑制剂奠定了基础。