FIBR: Planning: Proteomic Analysis of Critical Targets of Environmental Stress

这项拟议的工作将发现生物体中的热“薄弱环节”。也就是说，当生物体遭遇越来越高的温度时，哪些蛋白质最先受到温度的破坏？为了做到这一点，这项工作将利用一种分子伴侣，一种识别并结合受损蛋白质的特殊蛋白质。这些伴侣将被分离--与它们结合的受损蛋白质也将被分离。受损的蛋白质将通过肽质量指纹图谱进行识别，这是一种新的、最先进的技术。这项拟议的工作是一个试点项目，将在模式细菌物种大肠杆菌的种群中寻找在不同温度下进化的薄弱环节，并测试低温种群中的薄弱环节是否在高温种群中得到加强。拟议的工作将为一种方法学奠定基础，通过这种方法，可以识别其基因组已被表征的任何有机体的薄弱环节，以及与任何自然或人为压力有关的薄弱环节。这些薄弱环节潜在地决定了生物体的生态和进化极限、它们可以达到的最大年龄、对疾病的易感性等--但到目前为止，还不可能确定这些薄弱环节本身。该项目也将是首批展示多肽质量指纹图谱对这一科学领域问题的适用性的项目之一。

The proposed work will discover thermal "weak links" in organisms. That is, as organisms encounter progressively higher temperatures, which are the first proteins to be damaged by temperature? To do so, the work will exploit a molecular chaperone, a special protein that recognizes and binds to damaged proteins. These chaperones will be isolated - and with them the damaged proteins that they bind. The damaged proteins in turn will be identified by peptide mass fingerprinting, a novel, state-of-the-art technique. The proposed work is a pilot project that will seek weak links in populations of a model bacterial species, E. coli, that have been evolving at different temperatures, and test whether the weak links in low-temperature populations have been strengthened in high temperature populations.The proposed work will set the stage for a methodology whereby weak links can be identified for any organism whose genome has been characterized, and in relation to any natural or anthropogenic stress. Potentially these weak links determine the ecological and evolutionary limits of organisms, the maximum age they can reach, susceptibility to disease, etc. - but heretofore it has not been possible to identify the weak links themselves. The project will also be among the first to demonstrate the applicability of peptide mass fingerprinting to problems in this area of science.