Phosphorylated Carbohydrate Transport Across the Bacterial Cell Envelope

磷酸化碳水化合物跨细菌细胞膜的运输

• 批准号: RGPIN-2018-06546
• 负责人: Moraes, Trevor
• 金额: $ 4.23万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714309
• 关键词: Phosphorylated; Carbohydrate; Transport; Across; Bacterial

Bacteria depend on the acquisition of a diverse set of nutrients from their hosts to engage in successful growth, colonization or pathogenesis. Commensal and/or pathogenic bacteria compete with host cells for substrates ranging from metals to vitamins. As this competition can influence the outcome of infection, pathogens have developed a variety of nutrient uptake mechanisms to increase their competitiveness and hence their ability to colonize and productively infect their hosts. The long-term objective of my research programs is to elucidate the molecular basis of bacterial nutrient uptake by isolating membrane proteins and performing atomic resolution structural and functional characterization using a unique combination of X-ray crystallography and other biophysical tools such as microscale thermophoresis and biolayer interferometry, surface plasmon resonance and isothermal titration calorimetry. My NSERC research focuses on nutrient uptake mechanisms via the binding-protein dependent transporters (BPDTs), which target diverse ligands such as metal ions, amino acids and sugars and are recognized as virulence determinants in many bacterial infections. My short-term objectives outlined in this proposal are to decipher the mechanism of AfuABC a binding protein dependent transporter that we determined is responsible for active uptake of phosphorylated carbohydrates. Our structural insights will provide the molecular basis of nutrient scavenging from the host and how these bacteria compete for nutrients in complex environments. Aim 1. Characterize the identify AfuABC homologs and paralogs for substrate specificity and transport. Aim 2. Dissect the mechanism of transport of phospho-sugars by AfuABC using crystal structures, growth assays and biomolecular interactions to probe the interaction between the AfuA and AfuBC components of the uptake system. Aim 3. Determine the role of transporters in the pathogenesis and the source of the substrates that are acquired by these bacteria. These aims will lead to a critical understanding of phospho-sugar nutrient acquisition by bacterial pathogens and how this process provides a competitive advantage in their environment. In addition, this research program will provide tools to investigate membrane proteins and their interactions, providing an ideal and unique training environment for my 3 undergraduates, 6 graduate students and 2 postdoctoral fellows, 3 of whom will be involved in conducting the microbiology, biochemistry and biophysical methods outline in aims 1 and 2. In keeping with my successful training of highly qualified personnel to date, my trainees will have the opportunity to contribute to high-impact publications and will gain broad expertise in membrane protein structural biology, microbiology, and biochemistry, preparing them for careers in basic scientific research and biotechnology.

细菌取决于从其宿主中获得多种营养物质以成功生长，定殖或发病机理。共生和/或致病细菌与宿主细胞竞争，从金属到维生素的底物。由于该竞争可以影响感染的结果，因此病原体已经开发出各种营养摄取机制，以提高其竞争力，从而实现殖民和有效地感染宿主的能力。我的研究计划的长期目标是通过隔离膜蛋白并使用X射线晶体学和其他生物物理工具的独特组合（例如微量粒料和biophophoresis and Biolayer biolayer bytredortry，表面上的浮球剂，iSotherations and patry）和Isotheration和Isotheration和Isotheration和Isotheralmothation和Isotheration和Isotheration antrane和Isotheration和Isotheration titr，通过隔离膜蛋白并进行原子分辨率结构和功能表征来阐明细菌养分摄取的分子基础。 我的NSERC研究重点是通过结合蛋白依赖性转运蛋白（BPDTS）的营养摄取机制，该转运蛋白（BPDTS）靶向多种配体，例如金属离子，氨基酸和糖，并被识别为许多细菌感染中的毒力决定因素。我在该提案中概述的我的短期目标是破译我们确定的结合蛋白依赖性转运蛋白的机制，我们确定该机理负责积极吸收磷酸化的碳水化合物。我们的结构见解将为宿主的营养清除以及这些细菌如何在复杂环境中竞争营养素的分子基础。 目的1。表征鉴定AFUABC同源物和旁系同源物的底物特异性和运输。 AIM 2。使用晶体结构，生长测定和生物分子相互作用来探究AFUABC将磷酸糖转运的机理，以探测摄取系统的AFUA与AFUBC成分之间的相互作用。 AIM 3。确定转运蛋白在这些细菌获得的发病机理和底物的来源中的作用。 这些目标将导致对细菌病原体获取磷酸糖营养素的批判性了解，以及该过程如何在其环境中提供竞争优势。此外，该研究计划将提供研究膜蛋白及其相互作用的工具，为我的三名本科生，6名研究生和2名博士后研究员提供理想和独特的培训环境，其中3名将参与微生物学，生物化学和生物物理学方法的培训，以促进1和2的培训。高影响力的出版物，并将在膜蛋白结构生物学，微生物学和生物化学方面获得广泛的专业知识，并为他们的职业做准备基础科学研究和生物技术。