Characterization and Design of Histone-Derived Antimicrobial Peptides

 描述(申请人提供)：对常规抗生素产生抗药性的细菌日益成为公共卫生关注的问题，抗菌肽(AMPs)是对抗这些细菌的潜在替代药物。一个耐人寻味的AMP家族是组蛋白衍生的抗菌肽(HDAP)。HDAP是从自然来源中分离出来的，其中许多多肽的机制已经通过实验确定，例如在我们之前资助的地区赠款期间进行的工作中。我们实验室的其他工作也设计了一系列基于组蛋白晶体结构的新型HDAP。然而，相对较少的工作集中在HDAP的合理优化上，以设计更具活性的多肽。本提案中的研究旨在通过考虑合理设计HDAP的系统方法来解决这一差距。在这个拟议的设计中，我们将重点关注三个因素的作用：碱性氨基酸的Arg组成，活性多肽的最小长度和杂交肽的产生。我们将结合细菌分析、共聚焦显微镜、光谱测量和分子动力学模拟来探索这些因素如何在分子水平上影响多肽活性。虽然已知这些因素会影响其他AMP的活性，但导致这些影响的结构-功能关系尚不清楚。因此，我们希望这项拟议工作的结果可以被推广，以深入了解这些因素如何影响其他AMP的活性。HDAP是用于这一目的的理想的多肽家族，因为它们通过不同的机制工作，一些通透膜和另一些转运到细胞内并与细胞内成分相互作用。在阐明了HDAP中这三个特征的趋势之后，我们将结合这些信息来创建一系列新的HDAP，以优化多肽活性的单个因素。因此，我们可以验证这是否是一种在HDAP中进行多肽设计的有用方法，也可以应用于其他多肽系统。总之，这些研究的洞察力将促进HDAP作为潜在治疗剂的发展，并提供对HDAP结构-功能关系的见解，这些关系可能会推广到其他阳离子AMP家族。除了这些科学目标外，这项拟议的研究还非常重视培训，因为这项工作将由几名本科生研究人员和几名应届毕业生进行，卫尔斯理学院是一所本科女子学院。

 DESCRIPTION (provided by applicant): Bacteria that have developed resistance to conventional antibiotics are an increasing public health concern, and antimicrobial peptides (AMPs) represent a potential alternative to combat these bacteria. One intriguing family of AMPs is the histone-derived antimicrobial peptides (HDAPs). HDAPs have been isolated from natural sources, and the mechanisms for many of these peptides have been determined experimentally, such as in work performed during our previously funded AREA grant. Other work in our lab has designed a series of novel HDAPs based on histone crystal structures. However, relatively little effort has focused on the rational optimization of HDAPs to engineer more active peptides. The research in this proposal aims to address this gap by considering a systematic approach for the rational design of HDAPs. In this proposed design, we will focus on the role of three factors: the Arg composition of basic amino acids, the minimal length of active peptides and the creation of hybrid peptides. We will use a combination of bacterial assays, confocal microscopy, spectroscopic measurements, and molecular dynamics simulations to probe how each of these factors influences peptide activity on a molecular level. Although these factors are known to influence the activity of other AMPs, the structure-function relationships leading to these effects are not well understood. Thus, we expect that results from this proposed work can be generalized to give insight into the how these factors influence the activity of other AMPs. HDAPs are an ideal family of peptides to use for this purpose since they operate through different mechanisms, with some permeabilizing membranes and others translocating into cells and interacting with intracellular components. After elucidating trends for these three characteristics in HDAPs, we will combine this information to create a novel series of HDAPs that optimize the individual factors for peptide activity. We can thus validate whether this is a useful approach to peptide design in HDAPs that could be applied to other peptide systems. Together, the insight from these studies will both promote the development of HDAPs as potential therapeutic agents and provide insights into HDAP structure-function relationships that potentially can be generalized to other families of cationic AMPs. In addition to these scientific goals, this proposed research also has a strong emphasis on training as the work will be carried out by several undergraduate researchers and a few recent graduates at Wellesley College, an undergraduate women's college.