The aim of this project is to develop chemical methods and apply these to specific biological areas. Two problems of current interest are 1) chemistry of glutathione and its adducts with epoxides; 2) mechanisms of separation and purification of polypeptides by high-performance liquid chromatography. The level of success of a program on bio-mechanisms, such the one posed by the glutathione pathway, is largely dependent on the ability to generate the chemical information necessary for the interpretation of biological experiments. A systematic program was initiated and eventually has culminated in the development of unequivocal syntheses of the requisite compounds, methods of separation for diastereoisomeric conjugates of glutathione with alkene and arene oxides, establishment of the absolute configuration of these thioether stereoisomers, and correlation of synthetic compounds with enzymatically produced conjugates. The high-performance liquid chromatography analysis of peptides has focused on the role of hydrophobicity in the separation mechanisms by reveral phase liquid chromatography. Experiments have been designed and implmented which alter the intrinsic hydrophobicity of peptide molecules during chromatographic analysis. These changes are introduced in a predictable fashion such that valuable structural information is derived from conventional reversed-phase experiments. Successful applications of this approach include: characterization of amphibian peptides in small-cell carcinoma, purification of a protein seemingly synthesized in response to estrogen exposure, and characterization of two forms of epidermal growth factor isolated from mouse submaxillary gland.
