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Characterization of the ferrous iron transporter Feo

亚铁转运蛋白 Feo 的表征

基本信息

批准号：
10053292
负责人：
Shelley M. Payne
金额：
$ 37.97万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-03 至 2022-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10053292
关键词：
ATP phosphohydrolase Active Biological Transport Animal Model C-terminal Cell membrane Cells Cholera Complex Cytoplasmic Tail Diarrhea Disease Elements Energy-Generating Resources Environment GTP Binding GTP-Binding Proteins Gastrointestinal tract structure Goals Grant Guanosine Triphosphate Phosphohydrolases Habitats Human Hydrolysis In Vitro Intestines Iron Knowledge Life Membrane Modeling Mutation Analysis N-terminal Nature Nutrient Organism Oxygen Pathogenicity Politics Proteins Proton-Motive Force Reagent Role Site Small Intestines Source Structural Models Structure System Testing Transport Process Vibrio cholerae Virulence Work diarrheal disease experience experimental study health economics host colonization human pathogen in vivo member novel organizational structure pathogen periplasm protein B protein protein interaction response stoichiometry tool uptake

项目摘要

The diarrheal disease cholera poses enormous health, economic, and political burdens worldwide. The causative agent of cholera, Vibrio cholerae, is endemic in aquatic habitats, but it is capable of infecting the human small intestine, causing a massive, life-threatening diarrheal response. To colonize the host and cause disease, V. cholerae must acquire essential nutrients, including iron, in the host environment. The predominant form of iron present in the small intestine is ferrous iron. Thus, it is critical to understand the mechanisms of ferrous iron uptake in V. cholerae. The major ferrous iron transporter in V. cholerae is Feo. The Feo system is widely distributed among all bacterial species, and has important functions in the virulence of several pathogenic species; nevertheless, very little is known about its structure and mechanism of transport. In V. cholerae, the Feo transporter is composed of three proteins, FeoA, FeoB, and FeoC. The membrane-embedded C-terminal domain of FeoB is likely to form the pore for iron transport, and, interestingly, its N-terminal domain has homology to small eukaryotic G proteins, suggesting a novel mechanism of transport. The roles of FeoA and FeoC are unknown. We recently demonstrated that the three Feo proteins associate to form a higher order complex in vivo. This represents the first structural analysis of the mature, membrane- embedded, active Feo complex in vivo. The goal of this proposal is to determine the overall structure of this large complex in order to build and test models for the mechanism of iron transport. In our first specific aim, we will determine the mass and stoichiometry of the native Feo complex. This will lay the groundwork for a structural model of the active Feo transporter. In aim 2, we will refine this model by delineating the sites of interaction within and between the members of the complex. We will then test the functional significance of these interactions for complex formation and iron transport activity. In aim 3, we will determine the source of energy for transport through Feo. These studies will significantly advance our knowledge of the structure and function of this important and unique iron transporter. Significantly, all our studies will be carried out using active, membrane-associated Feo complexes in vivo, giving our results an undeniable relevance over the in vitro studies that currently dominate the Feo field. As our previous work shows, V. cholerae is an ideal model organism for the study of Feo. We have already assembled most of the strains and reagents needed, and we, and our collaborators, have the expertise required to carry out the proposed experiments.

霍乱造成了巨大的健康、经济和政治负担国际吧霍乱的病原体，霍乱弧菌，是地方性的水生栖息地，但它能够感染人的小肠，导致严重的危及生命的腹泻反应为了定殖宿主并引起疾病，霍乱弧菌必须获得必需的营养，包括铁在内的有害物质。铁的主要形式存在于小肠是亚铁。因此，了解二价铁吸收的机制至关重要霍乱弧菌。霍乱弧菌中主要的亚铁转运蛋白是FeO。Feo系统是广泛分布于所有细菌种，并在致病性中具有重要作用。几种致病物种;然而，对其结构和机制知之甚少交通。在霍乱弧菌中，FeO转运蛋白由三种蛋白质组成，FeoA，FeoB，和FeoC。FeoB的膜包埋的C-末端结构域可能形成孔，铁转运蛋白，有趣的是，其N-末端结构域与真核小G 蛋白质，这表明了一种新的运输机制。FeoA和FeoC的作用是未知我们最近证明，这三种FeO蛋白结合形成一种更高的体内有序复合体。这是第一次对成熟的膜结构进行分析，包埋的活性FeO复合物。本提案的目标是确定总体这一大型复杂的结构，以建立和测试模型的机制铁运输在我们的第一个具体目标，我们将确定质量和化学计量的天然 Feo复合物。这将奠定基础的结构模型的积极FeO运输。在目标2中，我们将通过描述复杂的成员。然后，我们将测试这些相互作用的功能意义，复合物形成和铁转运活性。在目标3中，我们将确定能量的来源通过Feo运输。这些研究将大大提高我们对这一重要而独特的铁转运蛋白的结构和功能。值得注意的是，我们所有的研究将进行使用活性，膜相关的FeO复合物在体内，给我们的结果与目前主导Feo领域的体外研究具有不可否认的相关性。作为我们前期工作表明，霍乱弧菌是研究FeO的理想模式生物。我们有已经组装了所需的大部分菌株和试剂，我们和我们的合作者，具备开展拟议实验所需的专业知识。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Vibrio cholerae CsrA Regulates ToxR Levels in Response to Amino Acids and Is Essential for Virulence.

DOI：
10.1128/mbio.01064-15
发表时间：
2015-08-04
期刊：
mBio
影响因子：
6.4
作者：
Mey AR;Butz HA;Payne SM
通讯作者：
Payne SM

Iron Transport and Metabolism in Escherichia, Shigella, and Salmonella.

DOI：
10.1128/ecosalplus.esp-0034-2020
发表时间：
2021-12-15
期刊：
EcoSal Plus
影响因子：
0
作者：
Mey AR;Gómez-Garzón C;Payne SM
通讯作者：
Payne SM

The Vibrio cholerae VctPDGC system transports catechol siderophores and a siderophore-free iron ligand.

DOI：
10.1111/j.1365-2958.2011.07775.x
发表时间：
2011-09
期刊：
Molecular microbiology
影响因子：
3.6
作者：
Wyckoff EE;Payne SM
通讯作者：
Payne SM

Developing Colorimetric and Luminescence-Based High-Throughput Screening Platforms for Monitoring the GTPase Activity of Ferrous Iron Transport Protein B (FeoB).

开发基于比色和发光的高通量筛选平台，用于监测亚铁转运蛋白 B (FeoB) 的 GTP 酶活性。

DOI：
10.1177/2472555219844572
发表时间：
2019
期刊：
SLAS discovery : advancing life sciences R & D
影响因子：
0
作者：
Veloria,John;Shin,Minhye;Devkota,AshwiniK;Payne,ShelleyM;Cho,EunJeong;Dalby,KevinN
通讯作者：
Dalby,KevinN

Vanadate inhibits Feo-mediated iron transport in Vibrio cholerae.

钒酸盐抑制霍乱弧菌中 Feo 介导的铁转运。