Bridging molecular studies of model and real biological systems with biophysics experiments on complex samples

将模型和真实生物系统的分子研究与复杂样品的生物物理学实验联系起来

• 批准号: RGPIN-2018-05154
• 负责人: Booth, Valerie
• 金额: $ 2.62万
• 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: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=645811
• 关键词: Bridging; molecular; studies; model; real

The theme of the NSERC-based research in our lab is bridging model and real biological systems. The rationale for this approach is that detailed, molecular level, structural and dynamics research on biological molecules is generally done with systems of very simple composition, e.g. with one type of lipid and one type of protein. However, it's not always clear how the results from these simple systems compare to what goes on in the real, much more complex biological systems. Thus, our approach is to build bridges between the biophysical methods that have been developed for simple systems and extend them to much more complex biological systems in order to gain important insights into real life molecular structure and dynamics. ******With this Discovery Grant renewal, I am planning two main foci. Objective 1 will build on our already successful NSERC research into how antimicrobial peptides (AMPs) interact with intact bacteria. AMPs are a crucial component of the innate immune system of many organisms and can protect against a variety of invading pathogens including bacteria, viruses, and microbial eukaryotes. However, our understanding of how AMPs work does not extend much past how they interact with one component of pathogens: the lipids. With Objective 2, we will apply our expertise in performing biophysics with complex biological samples into a new area to us, soft interactions in molecular crowding, initially as applied to Intrinsically Disordered Proteins (IDPs). Molecular crowding, i.e. the effect of the very high concentration of biomolecules inside cells, is important because it can modify the structure and function of biological molecules compared to what is observed in dilute conditions via traditional biochemistry experiments.******The potential impact of Objective 1 relates to the enormous disparity between conditions used for studies of AMP molecular mechanisms, versus experiments where biological functional is actually observed; it takes an AMP to lipid ratio of ~10,000 greater to see inhibition of cell growth as compared to the ratio needed to see lipid bilayer disruption with a model lipid system. Thus, it is of high interest to find out with which components, besides the lipids, the AMP is interacting and how the non-lipid interactions modify the AMP-induced membrane disruption. There has been a high level of interest in the earlier phase of our work in this area, as evidenced by three invited reviews on the topic, and we expect this high interest to continue.******Our results from Objective 2 will impact the field via building a much needed and novel bridge to better understand how the large body of work performed on simple crowding systems can be applied to real cells. Conversely, our results should also help explain observations gleaned from in-cell experiments. Additionally, our work will provide valuable insights into parameters of interest in getting quality data with any kind of protein via in-cell NMR.**

本实验室基于NSERC的研究主题是桥接模型和真实的生物系统。这种方法的基本原理是，对生物分子的详细的分子水平、结构和动力学研究通常是用组成非常简单的系统进行的，例如，用一种类型的脂质和一种类型的蛋白质。然而，我们并不总是清楚这些简单系统的结果如何与真实的、复杂得多的生物系统的结果进行比较。因此，我们的方法是建立桥梁之间的生物物理方法，已经开发了简单的系统，并将其扩展到更复杂的生物系统，以获得重要的见解到真实的生活分子结构和动力学。* * 目标1将建立在我们已经成功的NSERC研究抗菌肽（AMP）如何与完整的细菌相互作用。 AMP是许多生物体的先天免疫系统的重要组成部分，并且可以保护免受各种入侵病原体，包括细菌，病毒和微生物真核生物。 然而，我们对AMP如何工作的理解并没有超出它们如何与病原体的一种成分：脂质相互作用。在目标2中，我们将把我们在复杂生物样品的生物物理学方面的专业知识应用到一个新的领域，即分子拥挤中的软相互作用，最初应用于内含子无序蛋白质（IDP）。分子拥挤（molecular crowding），即细胞内生物分子浓度非常高的效应，是很重要的，因为它可以改变生物分子的结构和功能，而不是通过传统的生物化学实验在稀释条件下观察到的。目标1的潜在影响涉及AMP分子机制研究所用条件与实际观察到生物功能的实验之间的巨大差异;与模型脂质系统观察到脂质双层破坏所需的比率相比，AMP与脂质的比率需要高约10，000才能观察到细胞生长抑制。 因此，它是高度感兴趣的，以找出哪些组件，除了脂质，AMP是相互作用和非脂质相互作用如何修改AMP诱导的膜破坏。对我们在这一领域工作的早期阶段表现出很高的兴趣，就这一专题邀请进行的三次审查就证明了这一点，我们希望这种高度兴趣将继续下去。我们的目标2的结果将通过建立一个急需的和新颖的桥梁来影响该领域，以更好地理解如何将在简单拥挤系统上进行的大量工作应用于真实的细胞。相反，我们的结果也应该有助于解释从细胞内实验中收集到的观察结果。此外，我们的工作将为通过细胞内NMR获得任何类型蛋白质的质量数据提供有价值的见解。