13] Synthesis of oligopeptides for the study of cyclic nucleotide-dependent protein kinases

Abstract

Publisher Summary This chapter discusses methods used in solid-phase synthesis of peptides and to the use of peptides to assay cyclic nucleotide-dependent protein kinases. It describes the strategies for peptide synthesis that are applicable to commercially available reagents and equipment, specifically the use of chloromethylated polystyrene resin as solid support, the Boc group for α -NH2 protection, and N , N' -dicyclohexylcarbodiimide (DCC) as a coupling reagent. Although several innovative uses have been made of synthetic peptides in the study of cyclic nucleotide-dependent protein kinases, their most common application has been as substrates to study the determinants of specificity of these enzymes. In addition to studies of substrate specificity, peptide substrates modeled after sequences in intact proteins are useful in delineating substrate directed, rather than enzyme-directed, influences of effector molecules. In addition, the potential exists for the development of modified synthetic peptides, which would serve as irreversible inhibitors or would be suitable for affinity or photoaffinity labeling of the active sites of these enzymes.