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

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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.