Regulation mechanisms of ABC transporters

ABC转运蛋白的调控机制

• 批准号: 10260611
• 负责人: Heather Wendy Pinkett
• 金额: $ 31.07万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10260611
• 关键词: ATP-Binding Cassette Transporters; Adenosine Triphosphate; Antibiotics; Bacteria; Binding; Binding Proteins; Biochemical; Biochemistry; Biological Assay; Biological Models; Biophysics; Cell Survival; Cell membrane; Cells; Classification Scheme; Complex; Core Assembly; Cues; Cytolysis; Cytoplasm; Data; Development; Ensure; Environment; Epithelial Cells; Family; Gatekeeping; Genes; Goals; Heme; Hydrolysis; Immune; Immune response; Infection; Iron; Laboratories; Length; Mediating; Membrane; Molecular; Molecular Conformation; Nickel; Nutrient; Organism; Peptides; Play; Positioning Attribute; Proteins; Recycling; Regulation; Research; Research Personnel; Resistance development; Rice; Role; Site; Specificity; Structure; Sulfur; System; Testing; Toxic effect; Toxin; Virulence; Work; antimicrobial drug; antimicrobial peptide; base; cofactor; dimer; experimental study; insight; microbial; novel; pathogen; pathogenic bacteria; programs; recruit; sensor; structural biology; uptake

PROJECT SUMMARY This proposal seeks to understand how proteins located in the cell membrane work as gatekeepers to selectively allow compounds into or out of the cell. Such gatekeepers are known as or ATP-binding cassette (ABC) transporters, because they use the energy of ATP (adenosine triphosphate) hydrolysis to transport compounds across the cell membrane. Bacterial ABC importers are essential for organism survival, controlling the rate of uptake for nutrients scavenged from the bacterium's environment. Control of the rate of transport precludes over-accumulation of a nutrient that is beneficial at low concentrations, but is potentially toxic at high concentrations. While a subset of ABC proteins contain an additional “accessory” domain that can regulate the uptake of compounds by shutting off the transporter, it is unclear why certain transporters contain these domains while others do not. However, we do understand that certain transporters are “turned off” when a specific compound or protein binds to this accessory domain. Other accessory domains regulate by “sensing” changes in the microenvironment and reacting accordingly. To decipher this mechanism of regulation, the PI's laboratory combines biochemical and biophysical experiments with structural biology to understand how these accessory domains play a role in transport regulation, which in restricts or allows nutrients to enter the cell. This research program will define the molecular mechanism that controls nutrient uptake and allow researchers to understand how multiple transport systems work in concert within an organism to maintain cell survival. We will test our hypothesis that regulation of transporter activation via a sensing accessory protein. The proposed research will decipher the complex circuitry of regulation in a model system in three Aims to: (1) understand how PepT SBPs select for different substrates within the microenvironment (i.e., nutrients, cofactors and peptides); (2) determine how the assembly of the core transporter dictates transport selectivity and efficiency (3) reveal how PepT transporters regulate the import of substrates into the cell through the activation of a novel regulatory domain. This research program has set out to close critical gaps in the understanding of the fundamentals of the transport mechanism present in all bacteria. The results will yield insights into how regulatory domains modulate transport across all organisms, crucial for cell viability.

项目摘要 该建议旨在了解蛋白质如何位于细胞膜工作中，作为看门人 有选择地允许化合物进入或退出细胞。这样的网守被称为或ATP结合 纸盒（ABC）转运蛋白，因为它们使用ATP（三磷酸腺苷）水解的能量 在细胞膜上传输化合物。细菌ABC进口商对于 生物生存，控制从细菌清除营养的摄取率 环境。控制运输速率的控制排除了养分的过度积累 在低浓度下有益，但在高浓度下可能有毒性。而ABC的子集 蛋白质包含一个附加的“附件”域，可以调节化合物的摄取 关闭转运蛋白，尚不清楚为什么某些转运蛋白包含这些域而其他域 不要。但是，我们确实知道某些转运蛋白在特定时被“关闭” 化合物或蛋白质与该附件结构域结合。其他附件域通过“传感”来调节 微环境的变化并做出相应的反应。为了破译这种调节机制， PI的实验室将生化和生物物理实验与结构生物学结合在一起 了解这些附属域如何在运输法规中发挥作用，该法规在限制或 允许营养进入细胞。该研究计划将定义分子机制 控制营养的吸收，并使研究人员能够了解多个运输系统的工作方式 在生物体中的音乐会以维持细胞生存。我们将检验我们的假设 通过灵敏度辅助蛋白激活转运蛋白。拟议的研究将破译 模型系统中调节的复杂电路三个目的：（1）了解pept sbps如何 选择微环境中的不同底物（即营养，辅助因子和胡椒粉）； （2） 确定核心转运蛋白的组装如何决定运输选择性和效率（3） 揭示pept转运蛋白如何通过激活A的激活调节底物进口到细胞中 新型监管领域。该研究计划已着手弥合理解的关键差距 所有广播中存在的运输机制的基本原理。结果将产生见解 调节结构域如何调节所有生物体的运输，对细胞生存能力至关重要。