The principal aim of the previous grant was to use a combination of biophysical and computational techniques to evaluate the key features in the folding/misfolding of the 17 KD beta protein interleukin-1beta (IL-1beta). We have established that the slow folding of this beta-trefoil protein is largely controlled by "topological frustration". During this time period we also became interested in understanding the role conformational dynamics plays in the biological function of this family of proteins. The change in the Title of the grant from "Investigation of the Folding Mechanism of a Beta Protein" to "Investigation of the Functional/Folding Landscapes of the IL-1 Family" reflect this change in research focus. In addition, we have added Dr. Jose Onuchic as a co-Pi on this proposal we have established an integrated experimental/theoretical approach towards the questions we wish to address. It is apparent that the regulation of the processing of IL-1beta is of central importance in bacterial infection and septic shock;autoimmune diseases and some cancers1-8. Yet, little is known about the molecular requirements for processing and maturation of IL-1beta. The goals of the research outlined in this proposal are directed towards (a) investigating the interplay of the functional/folding landscapes within this family of proteins (b) understanding the structural basis for cleavage of the 31 KD precursor protein;(c) the structural characterization of the 31 KD precursor protein and the 24 KD partially cleaved protein;(d) understanding the role of the presequences in destabilizing the fold of the mature 17 KD protein. In summary, our goals are to look at the functional landscapes for related proteins, to develop methods for understanding the receptor binding and signal transmission by analyzing the energy landscapes for the cytokine agonist and antagonist receptor complexes, and exploring the conformational changes that accompany the processing of prolL-1beta from the inactive partially structured state to the active, mature IL-1beta. These studies are of both fundamental importance in understanding protein recognition and activity as well as of fundamental biological importance in understanding the conformational processes that regulate the activation of prolL-1beta.