Direct observation of the structure and function of cooperatively assembled membrane spanning pores formed by antimicrobial peptides.

项目简介：抗菌素耐药性的挑战激发了人们对了解先天免疫系统抗菌肽(AMPs)机制的兴趣。AMP通常形成跨膜孔作为其杀菌活性的一部分，然而，其组装和功能的确切机制仍然知之甚少。这在一定程度上是由于将功能模型膜系统与它们的询问技术相结合的挑战。本项目将结合应用于模型膜的多种单分子成像技术[1]来研究已有的和新的AMP的多肽相互作用的机制。通过单分子水平的研究，我们可以深入了解膜跨孔组装的机制。通过这种方法，您将通过定量三色单分子成像来研究Mainin-II/PGLa的协同活性，以了解多肽-肽相互作用的本质[2]，并量化组装的孔的活动动力学[3]。您还将把这些技术扩展到团队开发的新型AMP[4]。了解这种孔的作用机制将提供对分子自组装的基本理解，并确定孔本身的结构-功能关系，这是设计新型AMP的基础。被测试的多肽对癌细胞具有细胞毒活性，因此这项研究可能为下一代治疗和药物输送系统创造机会。一旦在模型膜中实现了多肽的表征，您将有机会在二维和三维细胞模型中研究它们的效果。研究环境：您将在物理-生命科学接口的多学科团队中工作，从生物化学、单分子技术、膜合成生物学和细胞生物学获得跨学科的经验。您将以多肽和蛋白质合成(JIN)和单分子膜成像(Castell)的旋转项目启动研究，为研究项目提供基础。监督团队在监督和出版方面有着出色的记录，并拥有独特的专业知识组合，将使您和项目受益，例如人造膜和单分子方法(Castell)、蛋白质/多肽生物化学(JIN)、AMPS和细胞生物学(Jones)以及组织模型和生物成像(Watson)。我们的应聘者来自热情的科学家，他们在一些概述的领域(生物化学|蛋白质/多肽处理|生物物理|物理化学|荧光)具有适当的知识/经验/想法，渴望跨学科学习并在候选人新的领域发展。我们并不期望在所有领域都有经验。我们鼓励所有背景的有才华和热情的应聘者申请。我们认识到，团队内部的多样性提供了更广泛的经验、视角和想法，可以在解决问题和为决策提供信息时参考。

Project Description:The challenge of antimicrobial resistance has ignited interest in understanding the mechanisms of antimicrobial peptides (AMPs) of the innate immune system. AMPs generally form membrane-spanning pores as part of their bactericidal activity, however, the precise mechanisms of assembly and function remain poorly understood. This is in part due to the challenges of interfacing functional model membrane systems with techniques for their interrogation.This project will combine multiple single molecule imaging techniques applied to model membranes [1] to study the mechanisms of peptide interaction of established and novel AMPs. By studying at the single molecule level we can gain otherwise inaccessible insight into the mechanisms of assembly of membrane spanning pores.With this approach you will study the synergistic activity of Magainin-II/PGLa by quantitative three-colour single-molecule imaging to understand the nature of peptide-peptide interactions [2] in pore formation, and quantify the activity dynamics of assembled pores [3]. You will also extend these techniques to novel AMPs developed by the team [4]. Understanding the mechanism of action of such pores will provide fundamental understanding into molecular self-assembly, alongside determining structure-function relationships of the pore itself, fundamental to design novel AMPs. The tested peptides possess cytotoxic activity against cancer cells, so this research may create opportunities for next generation therapeutics and drug delivery systems. Once peptide characterisation is achieved in model membranes you will have the opportunity to study their effect in 2-and-3D cell models.Research Environment: You will work within a multidisciplinary team at the physical-life science interface, gaining experience across disciplines from biochemistry, single molecule techniques, membrane synthetic biology and cell biology. You will initiate the research with rotational projects of peptide and protein synthesis (Jin), and single molecule membrane imaging (Castell), providing a foundation for the research project. The supervisory team have an excellent track record in supervision and publication, alongside a unique combination of expertise from which you and the project will benefit, - e.g. artificial membranes and single molecule methods (Castell), protein/peptide biochemistry (Jin), AMPs and cell biology (Jones) and tissue model and biological imaging (Watson). We applications from enthusiastic scientists with appropriate knowledge/experience/ideas in some of the areas outlined (biochemistry|protein/peptide handling|biological physics|physical chemistry|fluorescence) with an eagerness to learn across disciplines and develop in areas new to the candidate. We do not expect prior experience in all areas. Talented and enthusiastic candidates from all backgrounds are encouraged to apply. We recognise that diversity within a team provides a greater range of experience, perspectives and ideas to draw upon when tackling problems and informing decisions.