Bioinorganic Explorations of Host-defense Proteins

宿主防御蛋白的生物无机探索

基本信息

批准号：
10662538
负责人：
ELIZABETH M NOLAN
金额：
$ 31.44万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-15 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10662538
关键词：
Address Anti-Bacterial Agents Area Bacterial Physiology Binding Biochemical Biological Biophysics Case Study Cell physiology Chemistry Clinical Research Complex Development Diagnostic Disease Disulfides Environment Epithelial Cells Extracellular Space Funding Goals Growth Homeostasis Host Defense Human Human Pathology Immune Infection Infection prevention Inflammation Inflammatory Innate Immune Response Innate Immune System Intestines Invaded Investigation Ions Laboratories Leukocyte L1 Antigen Complex Link Metals Methionine Modeling Molecular Mus Nutrient Nutrient availability Nutritional Immunity Organism Outcome Pathology Peptide Hydrolases Physiology Play Post-Translational Protein Processing Property Proteins Research Resistance Role Rosaniline Dyes S100A8 gene S100A9 gene Site Starvation Structure Testing Therapeutic Transition Elements Urinary tract infection Uropathogenic E. coli Work biophysical properties design disulfide bond expectation extracellular host-microbe interactions human disease in vivo insight microbial molecular modeling neutrophil novel diagnostics oxidation pathogen pathogenic Escherichia coli pathogenic bacteria pathogenic fungus pathogenic microbe prevent response uptake

项目摘要

PROJECT SUMMARY The overarching goal of this renewal application is to elucidate the molecular basis for how human calprotectin (CP, S100A8/S100A9 oligomer) functions in the metal-withholding innate immune response, and to evaluate its impact on the physiology of uropathogenic Escherichia coli (UPEC), which cause the majority of urinary tract infections in humans. Transition metals are essential nutrients for all organisms, and the availability of these nutrients plays a critical role during microbial infection. Consequently, the human innate immune system launches a metal-withholding response and deploys metal-sequestering host-defense proteins into the extracellular space to limit metal availability and hinder pathogen growth. CP is an abundant and functionally versatile metal-withholding protein; it sequesters multiple metal nutrients including Mn(II), Fe(II), Ni(II) and Zn(II). Recent studies by our laboratory and others provide compelling evidence that the molecular speciation of extracellular CP is a heterogenous ensemble of different species that arises from different metal-bound forms as well as oxidative post-translational modifications. We hypothesize that this complex molecular speciation of CP, including the occurrence of methionine oxidation and disulfide bonding, has profound consequences for its extracellular function and lifetime. Recent studies by our laboratory and others also demonstrate that CP is a Cu-withholding protein. We hypothesize that CP sequesters both Cu(II) and Cu(I) and that this function impacts the physiology and metal homeostasis in diverse bacterial pathogens including UPEC. In Aim 1, we will examine disulfide bond formation within and between CP heterodimers, the biophysical properties of these disulfide-linked species, and their ability to sequester metals from bacterial pathogens. In Aim 2, we will evaluate the Cu(II/I)-binding properties of CP and the consequences of multi- metal sequestration by CP on UPEC as a case study. We expect that these investigations will advance the molecular model for how CP contributes to the metal-sequestering innate immune response, underscore the importance of considering CP species that result from oxidative posttranslational modification, and elucidate the molecular basis for Cu withholding by CP. Moreover, we expect that our studies of the interplay of CP and UPEC will provide new insight into how the host and pathogen compete for Cu and other nutrient metals. We further expect that the outcomes of this initiative may guide the design and development of novel diagnostic, preventative and therapeutic approaches for microbial infections and other pathologies such as inflammatory diseases that involve CP.

项目概要这一更新申请的总体目标是阐明人钙卫蛋白如何发挥作用的分子基础。（CP，S100A8/S100A9 寡聚物）在金属抑制先天免疫反应中发挥作用，并评估其对泌尿道致病性大肠杆菌 (UPEC) 生理学的影响，该大肠杆菌导致大多数尿路感染人类感染。过渡金属是所有生物体必需的营养物质，并且这些物质的可用性营养物质在微生物感染过程中起着至关重要的作用。因此，人类先天免疫系统启动金属扣留反应并将金属螯合宿主防御蛋白部署到细胞外空间限制金属可用性并阻碍病原体生长。 CP是一种丰富且功能丰富的多功能金属扣留蛋白；它可以隔离多种金属营养物，包括 Mn(II)、Fe(II)、Ni(II) 和锌(II)。我们实验室和其他人最近的研究提供了令人信服的证据，表明分子形态细胞外 CP 是不同物种的异质集合，由不同的金属结合产生形式以及氧化翻译后修饰。我们假设这种复杂的分子 CP的形态，包括蛋氨酸氧化和二硫键的发生，具有深远的影响对其细胞外功能和寿命的影响。我们实验室和其他人最近的研究也证明CP是一种Cu扣留蛋白。我们假设 CP 螯合 Cu(II) 和 Cu(I)，并且该功能影响多种细菌病原体的生理和金属稳态，包括统一经济共同体。在目标 1 中，我们将检查 CP 异二聚体内部和之间的二硫键形成，这些二硫键连接物种的生物物理特性，以及它们从细菌中螯合金属的能力病原体。在目标 2 中，我们将评估 CP 的 Cu(II/I) 结合特性以及多重影响的后果以 UPEC 上的 CP 金属封存为例进行研究。我们预计这些调查将推动 CP 如何促进金属螯合先天免疫反应的分子模型，强调了考虑氧化翻译后修饰产生的 CP 物种的重要性，并阐明 CP 扣留 Cu 的分子基础。此外，我们期望我们对 CP 和 CP 相互作用的研究 UPEC 将为宿主和病原体如何竞争铜和其他营养金属提供新的见解。我们进一步期望该倡议的成果可以指导新型诊断的设计和开发，微生物感染和其他病理（例如炎症）的预防和治疗方法涉及 CP 的疾病。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Biochemical and Spectroscopic Observation of Mn(II) Sequestration from Bacterial Mn(II) Transport Machinery by Calprotectin.

钙卫蛋白从细菌 Mn(II) 运输机械中螯合 Mn(II) 的生化和光谱观察。

DOI：
发表时间：
2018-01-10
期刊：
影响因子：
15
作者：
Hadley, Rose C;Gagnon, Derek M;Brophy, Megan Brunjes;Gu, Yu;Nakashige, Toshiki G;Britt, R David;Nolan, Elizabeth M
通讯作者：
Nolan, Elizabeth M

Calcium Binding to the Innate Immune Protein Human Calprotectin Revealed by Integrated Mass Spectrometry.

通过集成质谱法揭示钙与先天免疫蛋白人钙卫蛋白的结合。

DOI：
发表时间：
2020
期刊：
影响因子：
15
作者：
Adhikari, Jagat;Stephan, Jules R;Rempel, Don L;Nolan, Elizabeth M;Gross, Michael L
通讯作者：
Gross, Michael L

Biophysical Examination of the Calcium-Modulated Nickel-Binding Properties of Human Calprotectin Reveals Conformational Change in the EF-Hand Domains and His3Asp Site.

对人钙卫蛋白的钙调节镍结合特性的生物物理检查揭示了 EF-Hand 结构域和 His3Asp 位点的构象变化。

DOI：
发表时间：
2018-07-17
期刊：
影响因子：
2.9
作者：
Nakashige, Toshiki G;Bowman, Sarah E J;Zygiel, Emily M;Drennan, Catherine L;Nolan, Elizabeth M
通讯作者：
Nolan, Elizabeth M

Avian MRP126 Restricts Microbial Growth through Ca(II)-Dependent Zn(II) Sequestration.

禽类 MRP126 通过 Ca(II) 依赖性 Zn(II) 封存来限制微生物生长。