Nuclear Magnetic Resonance (NMR) studies of antimicrobial peptides in intact cells

完整细胞中抗菌肽的核磁共振 (NMR) 研究

• 批准号: 312676-2011
• 负责人: Booth, Valerie
• 金额: $ 2.91万
• 依托单位: Memorial University of Newfoundland
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=629992
• 关键词: Nuclear; Magnetic; Resonance; NMR; studies

Antimicrobial peptides (AMPs) are currently the subject of significant interest, in part due to their important role in our immune systems and in part due to their potential as novel therapeutics to combat the increasingly serious problem of microbial resistance to conventional antibiotics. However, there are gaps in both our understanding of how these molecules work and in our ability to use them as the basis for better treatments. One such gap results from the fact that the detailed mechanism of AMPs has only been studied in highly simplified systems that only approximate, at best, the much more complex cell envelopes against which the AMPs act. Microbe cell envelopes contain a number of additional components that likely affect how AMPs work and probably contribute to the ability of AMPs to kill microbes at levels that don't harm our own cells. We intend to bridge this gap by developing ways to use high resolution, quantitative techniques, like nuclear magnetic resonance (NMR), to investigate AMPs in whole, intact bacteria and mammalian cells. We will then compare our results to the results in the model systems to better understand how AMP actions are modified by the complexity present in real cells. We will also identify and characterize novel AMPs from cod. The cod genome represents an excellent source of potentially new and interesting antimicrobial peptide motifs, since cod have an underdeveloped adaptive immune system, but are literally swimming in a sea of pathogens and so presumably require especially efficacious AMPs. The anticipated significance of the work includes: supplying a badly needed connection between NMR studies that shed light on the mechanism of AMPs and those conditions under which activity is actually observed; developing new methods that enable the application of NMR to the study of any membrane protein in the context of whole cells; making available information of significance to the AMP community and those involved in cod and other fish breeding programs by identifying and characterizing novel marine AMPs.

抗微生物肽（AMP）目前是显著感兴趣的主题，部分是由于它们在我们的免疫系统中的重要作用，部分是由于它们作为新型治疗剂对抗日益严重的微生物对常规抗生素的耐药性问题的潜力。然而，在我们对这些分子如何工作的理解以及我们将它们作为更好治疗基础的能力方面都存在差距。一个这样的差距是由于这样的事实，即AMP的详细机制只在高度简化的系统中进行了研究，这些系统充其量只是近似AMP作用的更复杂的细胞包膜。微生物细胞包膜含有许多额外的成分，这些成分可能会影响AMP的工作方式，并可能有助于AMP在不伤害我们自己细胞的水平上杀死微生物的能力。我们打算通过开发使用高分辨率，定量技术，如核磁共振（NMR）来研究完整，完整的细菌和哺乳动物细胞中的AMP来弥合这一差距。然后，我们将比较我们的结果与模型系统中的结果，以更好地理解AMP行为如何被真实的细胞中存在的复杂性所修改。我们还将从鳕鱼中鉴定和表征新型AMP。鳕鱼基因组代表了潜在的新的和有趣的抗菌肽基序的一个很好的来源，因为鳕鱼具有不发达的适应性免疫系统，但实际上是在病原体的海洋中游泳，因此可能需要特别有效的AMP。这项工作的预期意义包括：在阐明AMP机制的NMR研究与实际观察到活性的条件之间提供迫切需要的联系;开发新方法，使NMR能够应用于整个细胞背景下的任何膜蛋白的研究;通过鉴定和表征新型海洋AMP，为AMP社区以及参与鳕鱼和其他鱼类养殖计划的人提供重要信息。