Molecular mechanisms of Gram-negative outer membrane organization

抗药性细菌感染频率的上升对人类健康构成了迫在眉睫的威胁。 试图产生新的抗生素来解决这个问题往往失败，因为外膜（OM） 革兰氏阴性细菌作为阻挡许多小分子进入的屏障。这个OM屏障是 与脂多糖（LPS）和外膜蛋白（OMPs）紧密结合， 在OM中不均匀。具体地说，已经证明外膜蛋白形成更高阶的组装体 在膜中称为“岛”。OM孔蛋白是三聚体和高度丰富的OMP，但尚不清楚如何 这种低聚状态影响它们的功能、插入OM或组装OMP岛。我 假设OMP三聚化对于OMP功能是关键的，并且从根本上是异源的基础， 组织OM。使用模型OMP，LamB，我开发了一套工具，使我能够 检验这个假设。首先，我将使用细菌遗传技术来确定OMP如何寡聚状态 影响OMP功能和LPS分配。其次，我将使用荧光显微镜技术， 表征OMP寡聚化在OMP岛组装中的作用。最后，我将确定如何本地 外膜蛋白的分泌影响外膜的组织结构。总之，我的工作将确定分子机制 管理OM的组织。重要的是，这将为破坏OM的战略提供信息， 设计OM渗透性抗生素。 这个奖学金的培训目标是继续增长我的科学知识，同时进一步发展 作为一个领导者和导师的技能，以实现我的职业目标，成为一个研究小组的领导者。作为一个 作为普林斯顿大学Silhavy实验室的一员，我可以发展一种教学理念， 在本科生和研究生的教学和指导方面。我会完善我的沟通技巧 通过大量的机会在各种会议上介绍我的工作。此外，分子生物学 普林斯顿大学的系是一个协作的、跨学科的环境， 发展思想和分享专门知识。总之，我相信，完成这一培训计划， 普林斯顿大学Silhavy实验室的研究环境将使我能够培养必要的技能， 管理我自己的研究小组

The rising frequency of drug-resistant bacterial infections pose an imminent threat to human health. Attempts to generate novel antibiotics to address this problem often fail because the outer membrane (OM) of gram-negative bacteria acts as a barrier that blocks the entry of many small molecules. This OM barrier is densely packed with lipopolysaccharides (LPS) and outer membrane proteins (OMPs) that organize heterogeneously within the OM. Specifically, it has been demonstrated that OMPs form higher-order assemblies in the membrane termed “islands”. The OM porins are trimeric and highly abundant OMPs but it is unknown how this oligomerization state influences their function, insertion into the OM, or the assembly of OMP islands. I hypothesize that OMP trimerization is critical for OMP function and fundamentally underlies the heterogenous organization of the OM. Using the model OMP, LamB, I have developed a set of tools that will allow me to examine this hypothesis. First, I will use bacterial genetic techniques to determine how OMP oligomeric state influences OMP function and partitioning of LPS. Second, I will use fluorescence microscopy techniques to characterize the role of OMP oligomerization in the OMP island assembly. Finally, I will determine how local secretion of OMPs affect OM organization. Together, my proposed work will define the molecular mechanisms that govern the organization of the OM. Importantly, this will inform strategies for disrupting the OM and in designing OM permeable antibiotics. The training goal of this fellowship is to continue to grow my scientific knowledge while further developing skills as a leader and mentor in order to fulfill my career goal of becoming a research group leader. Being a member of the Silhavy lab at Princeton University will allow me to develop a teaching philosophy and participate in the teaching and mentoring of undergraduate and graduate students. I will perfect my communication skills through ample opportunities to present my work at meetings and conferences. Additionally, the Molecular Biology Department at Princeton University is collaborative, interdisciplinary environment that promotes that development of ideas and sharing of expertise. Together, I believe that the completion of this training plan in the research environment of the Silhavy lab at Princeton University will allow me to develop the necessary skills to run my own research group.