Collaborative Research: Leveraging Biophysical Methods to Investigate the Interplay of Host Defense Mechanisms at Biological Membranes

合作研究：利用生物物理方法研究生物膜上宿主防御机制的相互作用

• 批准号: 1716608
• 负责人: Myriam Cotten
• 金额: $ 40.47万
• 依托单位: College of William and Mary
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1716608&HistoricalAwards=false
• 关键词: Collaborative; Research; Leveraging; Biophysical; Methods

Biological membranes and their constituents are involved in virtually all processes vital to living organisms, including the immune response. This research investigates how several agents of the immune response combine their antimicrobial effects by interacting with each other on bacterial cell membranes. The specific agents of interest are host defense peptides and copper ions. Host defense peptides are powerful molecules that act on a broad range of pathogenic cells, including bacteria, viruses, and fungi. Copper ions, which are present in immune cells, can damage the cell membranes of bacteria by oxidizing their phospholipids. Interestingly, some host-defense peptides contain a copper-binding motif that enables them to bind copper and become metallopeptides. In this project, the PIs will develop new bio-physical and biochemical tools to investigate how host defense metallopeptides and copper ions may synergize their antimicrobial effects. This project has the potential to produce new principles that could be useful to design novel antimicrobial biomaterials. In addition, the project will include activities that engage college and pre-college students from diverse backgrounds in hands-on science and enhance their analytical skills. The PIs will leverage the interdisciplinary and collaborative nature of their research, and passion for scientific research to spike scientific interest in young students and prepare high school students for a successful transition to college. The major goal of this research is a rigorous investigation of the interplay between host defense mechanisms at biological membranes. The molecular systems of interest are membrane-interacting host defense metallopeptides and other agents of the immune response that have the potential to unify their antimicrobial effects at lipid membranes. While host defense peptides have been studied for their ability to disrupt the conformational arrangement in bacterial cell membranes, the possibility that host defense peptides and other immune agents such as copper and oxidized lipids synergize their antimicrobial effects has not been explored on a molecular level. The PIs will test this hypothesis using peptides from fish (piscidins) and ticks (ixosins). They will leverage their expertise in the characterization of membrane assemblies and harness a platform of cutting-edge biophysical techniques (solid-state NMR, neutron diffraction, surface-sensitive methods) and biological assays to obtain structure-dynamics-function information on these metallopeptides under native-like conditions. The project will focus on model membranes to accurately and precisely control membrane composition and directly relate their molecular contents to specific functional and conformational behaviors of host defense metallopeptides and their interactions with bilayers. The project will feature assays on live cells relevant to the biological context of the chosen host defense peptides to investigate the contribution of lipid oxidation to the antimicrobial potency of the peptides, identify synergistic effects among peptides, and relate biological potencies to physicochemical properties. This project will generate multiple broad impact outcomes. First, the new findings and methodologies created will contribute to advancing scientific knowledge in the fields of membrane biophysics, structural biology, immunology, and biochemistry. These fundamental principles could be useful to design novel antimicrobial agents and biomaterials. Second, the scientific environment will foster partnerships between academia and national labs. Third, the outreach activities will stimulate the interest of college and pre-college students from diverse backgrounds in hands-on science, and enhance their analytical skills.

生物膜及其成分参与了几乎所有对生物体至关重要的过程，包括免疫反应。本研究调查了几种免疫应答剂如何通过在细菌细胞膜上相互作用来联合收割机组合它们的抗菌作用。感兴趣的特异性试剂是宿主防御肽和铜离子。宿主防御肽是作用于包括细菌、病毒和真菌在内的广泛致病细胞的强大分子。存在于免疫细胞中的铜离子可以通过氧化细菌的磷脂来破坏细菌的细胞膜。有趣的是，一些宿主防御肽含有铜结合基序，使它们能够结合铜并成为金属肽。在这个项目中，PI将开发新的生物物理和生物化学工具来研究宿主防御金属肽和铜离子如何协同其抗菌作用。该项目有可能产生新的原则，可能有助于设计新型抗菌生物材料。此外，该项目将包括让来自不同背景的大学生和大学预科生参与实践科学并提高他们的分析能力的活动。PI将利用其研究的跨学科和协作性质，以及对科学研究的热情，激发年轻学生的科学兴趣，并为高中生成功过渡到大学做好准备。 本研究的主要目标是严格调查生物膜上宿主防御机制之间的相互作用。感兴趣的分子系统是膜相互作用的宿主防御金属肽和免疫应答的其他试剂，其具有在脂质膜上统一其抗微生物作用的潜力。虽然已经研究了宿主防御肽破坏细菌细胞膜中构象排列的能力，但宿主防御肽和其他免疫剂如铜和氧化脂质协同其抗微生物作用的可能性尚未在分子水平上探索。PI将使用来自鱼（鱼杀菌素）和蜱（毒蜥毒素）的肽来检验这一假设。他们将利用他们在膜组件表征方面的专业知识，并利用尖端生物物理技术（固态NMR，中子衍射，表面敏感方法）和生物测定的平台，以获得这些金属肽在天然条件下的结构-动力学-功能信息。该项目将专注于模型膜，以准确和精确地控制膜组成，并直接将其分子含量与宿主防御金属肽的特定功能和构象行为及其与双层的相互作用联系起来。该项目将对与所选宿主防御肽的生物学背景相关的活细胞进行分析，以研究脂质氧化对肽的抗菌效力的贡献，确定肽之间的协同效应，并将生物学效力与理化性质联系起来。该项目将产生多个广泛的影响成果。首先，新的发现和方法将有助于推进膜生物物理学，结构生物学，免疫学和生物化学领域的科学知识。这些基本原理可用于设计新型抗菌剂和生物材料。第二，科学环境将促进学术界和国家实验室之间的伙伴关系。第三，外展活动将激发来自不同背景的大学生和大学预科生对动手科学的兴趣，并提高他们的分析能力。

项目成果

Coordination of Redox Ions within a Membrane-Binding Peptide: A Tale of Aromatic Rings

膜结合肽内氧化还原离子的配位：芳香环的故事

• DOI: 10.1021/acs.jpclett.1c00636
• 发表时间: 2021
• 期刊: The Journal of Physical Chemistry Letters
• 作者: Fu, Riqiang;Rooney, Mary T.;Zhang, Rongfu;Cotten, Myriam L.
• 通讯作者: Cotten, Myriam L.

Copper regulates the interactions of antimicrobial piscidin peptides from fish mast cells with formyl peptide receptors and heparin

• DOI: 10.1074/jbc.ra118.001904
• 发表时间: 2018-10-05
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kim, So Young;Zhang, Fuming;Cotten, Myriam L.
• 通讯作者: Cotten, Myriam L.

The host-defense peptide piscidin P1 reorganizes lipid domains in membranes and decreases activation energies in mechanosensitive ion channels

• DOI: 10.1074/jbc.ra119.010232
• 发表时间: 2019-12-06
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Comert, Fatih;Greenwood, Alexander;Mihailescu, Ella
• 通讯作者: Mihailescu, Ella

Enhancing the membrane activity of Piscidin 1 through peptide metallation and the presence of oxidized lipid species: Implications for the unification of host defense mechanisms at lipid membranes

通过肽金属化和氧化脂质种类的存在增强 Piscidin 1 的膜活性：对脂质膜宿主防御机制统一的影响

• DOI: 10.1016/j.bbamem.2020.183236
• 发表时间: 2020
• 期刊: Biochimica et Biophysica Acta (BBA
• 作者: Paredes, Steven D.;Kim, Sarah;Rooney, Mary T.;Greenwood, Alexander I.;Hristova, Kalina;Cotten, Myriam L.
• 通讯作者: Cotten, Myriam L.

Differential Interactions of Piscidins with Phospholipids and Lipopolysaccharides at Membrane Interfaces

• DOI: 10.1021/acs.langmuir.0c00017
• 发表时间: 2020-05-12
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Cetuk, Hannah;Maramba, Joseph;Sukharev, Sergei
• 通讯作者: Sukharev, Sergei