The objective of this research proposal is to investigate properties of chromatin-bound protease by proper design of synthetic inhibitors. This study will involve a dual program of organic synthesis with parallel biochemical studies. The design of inhibitors specific for chromatin-bound proteases will be very useful for studies on the role of the chromatin-bound proteases in gene function. The inhibitors that have been studied so far have a rather broad specificity and consequently are not useful in experiments with whole cells. Furthermore the synthesis of radioactive irreversible specific protease inhibitors will be useful for quantitation of chromatin-bound proteases in the cells at different stages of the cell cycle and development as well as in comparative studies on normal versus cancerous cells. The specific chemical class of potential inhibitors would involve the following: 1) To synthesize and design analogs of p-nitro-phenylacetate and phenylalanine as potential inhibitors of chromatin-bound protease. 2) The design of phenylalanine analogs which could be utilized in affinity chromatography for the purification of this protease. 3) Synthesis of radioactive irreversible specific protease inhibitors. The pursuit of these goals will be accomplished by collaboration with Dr. Chi-Bom Chae at the Department of Biochemistry, University of North Carolina.