Periplasmic Chaperone Network Organization and Mechanism

这个发现项目调查了在人类中发现的有益微生物的成熟过程。这些微生物有充满糖和脂肪的膜表面来包裹它们的细胞。蛋白质嵌入糖/脂肪混合物中，它们的适当结合对这些表面的完整性和功能至关重要。这个项目探讨了这三种分子是如何结合在一起的。将进行一系列系统的研究，以确定在组装过程中起关键作用的辅助蛋白的分子结构。该项目的第二部分将使用活细胞的图片来发现这些辅助蛋白在整个细胞中的表面分布。这些辅助蛋白在细胞中聚集在一起吗？还是均匀地洒在各处？此外，该项目还将通过为研究生和本科生提供研究机会，在研究生和本科生STEM教育中营造更具包容性的氛围，以及采用循证最佳实践进行教学，来扩大对科学的参与。本课题的目的是研究胞外伴体网络的组织和机制。这将通过两种一般方法来实现。首先是确定两种可溶性伴侣蛋白与未折叠的外膜蛋白复合物的溶液结构。这将通过整合正交、生物物理方法和计算建模的信息来完成。这项工作的结果将是新的结构，为这些伴侣蛋白如何通过维持溶液中未折叠的膜蛋白来确保有效的外膜蛋白生物生成提供关键信息。本项目的第二个基本工作是使用成像技术检查活细胞中关键伴侣蛋白的质周分布。后一项工作的主要问题是评估可溶性伴侣是否在这种缺乏任何外部能量来源的独特细胞室中混合良好。总之，这些结果将提供对细菌中膜蛋白生物发生的基本理解。这些信息可以用来推断真核生物线粒体同源系统的机制。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This discovery project investigates the maturation processes of beneficial microorganisms found in humans. These microbes have sugar- and fat-filled membrane surfaces that encapsulate their cells. Proteins are embedded within the sugar/fat mixture, and their proper incorporation is essential for the integrity and functions of these surfaces. This project addresses how these three types of molecules come together. A series of systematic investigations will be conducted to determine the molecular structures of helper proteins that play key roles in the assembly process. A second part of the project will use pictures of living cells to discover the surface distribution of these helper proteins throughout the cell. Are these helper proteins clumping all together in a cell? Or are they sprinkled evenly throughout? In addition, the PI will broaden the participation in science through research opportunities for graduate and undergraduate students, by nurturing a more inclusive climate in graduate and undergraduate STEM education, and by teaching using evidence-based best practices. The goals of this project are to investigate the periplasmic chaperone network organization and mechanism. This will be achieved using two general approaches. The first is to determine the solution structures of two of the soluble chaperone proteins in complex with an unfolded outer membrane protein. This will be accomplished by integrating information from orthogonal, biophysical methods and computational modeling. The outcome from this effort will be novel structures that provide key information on how these chaperones ensure efficient outer membrane protein biogenesis by maintaining unfolded membrane proteins in solution. The second fundamental effort addressed in this project is to examine the periplasmic distribution of a key chaperone in living cells using imaging techniques. The main question in this latter effort is to evaluate whether or not soluble chaperones are well mixed in this unique cellular compartment that lacks any external energy source. Together the outcomes will provide a fundamental understanding of membrane protein biogenesis in bacteria. This information can be used to infer mechanistic insights into homologous system in mitochondria of eukaryotes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.