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• 批准号: 10623041
• 负责人: Heather Wendy Pinkett
• 金额: $ 7.85万
• 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/10623041
• 关键词: ATP-Binding Cassette Transporters; Adenosine Triphosphate; Affinity; Antibiotic Resistance; Antibiotics; Award; Bacteria; Binding; Binding Proteins; Biochemical; Bioinformatics; Biological Assay; Biological Models; Biophysics; Carrier Proteins; Cell Survival; Cell membrane; Cells; Classification Scheme; Complex; Computer Models; Core Assembly; Coupled; Cues; Cytoplasm; DSPP gene; Data; Dipeptides; Docking; Electrophoretic Mobility Shift Assay; Ensure; Environment; Escherichia coli; Family; Funding; Gatekeeping; Genome; Glutathione; Goals; Haemophilus influenzae; Heme; Heme Iron; Hydrolysis; Immune; In Vitro; Inflammation; Laboratories; Membrane; Molecular; Nickel; Nontypable Haemophilus influenza; Nucleotides; Nutrient; Oligopeptides; Operon; Organism; Otitis Media; Parents; Pasteurellaceae; Peptide Transport; Peptides; Play; Prevalence; Production; Proteins; Regulation; Research; Research Personnel; Respiratory Disease; Respiratory System; Rice; Role; SH2D1A gene; Series; Source; Specificity; Starvation; Surface Plasmon Resonance; System; Testing; Therapeutic; Time; Tissues; Toxin; Transmembrane Domain; Virulence Factors; Work; World Health Organization; antimicrobial peptide; base; cofactor; comparative; experimental study; gene cloning; graduate student; heme a; heme-binding protein; host colonization; in silico; in vivo; insight; middle ear; mutant; novel; parent grant; pathogen; pathogenic bacteria; periplasm; programs; resistance mechanism; structural biology; uptake

I. SUMMARY OF THE FUNDED PARENT AWARD R01 GM140584 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 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. There are no changes to the funded parent award

一、资助家长奖 R01 GM140584 概要 该提案旨在了解位于细胞膜中的蛋白质如何充当守门人 选择性地允许化合物进入或离开细胞。这种看门人被称为 ATP 结合盒 (ABC) 转运蛋白，因为它们利用 ATP（三磷酸腺苷）水解的能量来转运化合物 穿过细胞膜。细菌 ABC 输入者对于生物体的生存至关重要，控制着细菌的传播速度 吸收从细菌环境中清除的营养物质。控制运输速度可防止过度运输 营养物质的积累，在低浓度下有益，但在高浓度下可能有毒。 虽然 ABC 蛋白的一个子集包含一个额外的“辅助”结构域，可以调节 通过关闭转运蛋白来合成化合物，目前尚不清楚为什么某些转运蛋白含有这些结构域，而 其他人则不然。然而，我们确实知道，当特定化合物出现时，某些转运蛋白会被“关闭” 或蛋白质结合到该辅助结构域。其他辅助域通过“感知”变化来调节 微环境并做出相应的反应。为了破译这种调节机制，PI 的实验室 将生物化学和生物物理实验与结构生物学相结合，以了解这些附件如何 结构域在运输调节中发挥作用，限制或允许营养物质进入细胞。这项研究 该计划将定义控制营养吸收的分子机制，并让研究人员了解 多个运输系统如何在生物体内协同工作以维持细胞存活。我们将测试我们的 通过传感辅助蛋白调节转运蛋白激活的假设。拟议的研究将 破译模型系统中复杂的调节电路，旨在实现以下三个目的：(1) 了解 PepT SBP 是如何工作的 选择微环境中的不同底物（即营养素、辅因子和肽）； (2)确定 核心转运蛋白的组装如何决定转运选择性和效率 (3) 揭示 PepT 如何发挥作用 转运蛋白通过激活新的调节域来调节底物进入细胞的过程。 该研究计划旨在弥合对交通基础知识理解的关键差距 机制存在于所有细菌中。结果将深入了解监管领域如何调节运输 在所有生物体中，对于细胞活力至关重要。 资助的家长奖励没有变化