SIMULTANEOUSLY SYNTHESIZED PEPTIDES ON CONTINUOUS CELLULOSE MEMBRANESAS SUBSTRATES FOR PROTEIN-KINASES

Sets of peptides with defined sequences, each on a separate spot, were synthesized simultaneously on continuous cellulose membranes (SPOTs(T M) membranes), which were originally designed for epitope studies. The applicability of the membrane-bound peptides as substrates for protei n kinases was tested using protein kinase A, protein kinase C and case in kinases I and II as model enzymes. We found that the peptide-membra ne complexes can serve as kinase substrates. Our results suggest that membrane-bound peptides offer a new potential for the investigation of substrate specificity of protein kinases. An advantage to this method is that there is not need for substrate identification and separation , which is required with high-volume random peptide libraries. Membran e-bound peptides may even form a basis for kinase assays with peptides lacking multiple basic amino acids, required for separation of the su bstrates in conventional assays. Problems connected with protein kinas e substrate specificity ca be investigated in any laboratory using the rapid and inexpensive SPOTs technique, as neither costly apparatus no r special experience in peptide synthesis is necessary.