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

这项拟议中的工作将发现生物体中的热“薄弱环节”。 也就是说，当生物体遇到逐渐升高的温度时，哪些蛋白质会首先被温度破坏？为了做到这一点，这项工作将利用分子伴侣，一种识别并结合受损蛋白质的特殊蛋白质。 这些分子伴侣将被分离出来，与它们结合的受损蛋白质也将被分离出来。 受损的蛋白质反过来将通过肽质量指纹图谱（一种新的最先进的技术）进行鉴定。 拟议的工作是一个试点项目，将寻找一个模型细菌物种，E。大肠杆菌，已经在不同的温度下进化，并测试是否在低温种群的薄弱环节已经加强了高温population.The拟议的工作将设置一个阶段的方法，从而可以确定薄弱环节的任何生物体的基因组已被定性，并与任何自然或人为的压力。 潜在地，这些薄弱环节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-但迄今为止，还不可能识别薄弱环节本身。 该项目也将是第一个证明肽质量指纹识别在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.