Pepsin Action on the New Synthetic Peptides 1. Pepsin action on benzyloxycarbonyl-glycyl-L-tyrosyl-L-phenylalanyl-glycine and its ethyl ester

[摘要] The synthesis is described of new pepsin substrates of benzyloxycarbonyl-glycyl-L-tyrosyl-L-phenylalanyl-glycine ethyl ester and benzyloxycarbonyl-glycyl-L-tyrosyl-L-phenylalanyl-glycine for studies on the specificity of pepsin, and thin layer chromatographic examination of the peptides prepared showed the new substrates are homogeneous and also, same examination of the incubation mixtures showed that two synthetic substrates are cleaved by pepsin at the L-tyrosyl-L-phenylalanyl bond and hydrolysis of these substrates by pepsin is achieved without transpeptidation. It is found that synthetic peptides are moderately soluble with the amount of the substrate up to a concentration of 0.7 mM in aqueous sodium citrate buffers (0.04 M) in the pH range 1.8-4.0, thus obviating the necessity for the adding of an organic solvent in the assay mixture. The kinetic parameters for synthetic substrates are tabulated in the following table. opt.temp. opt.pH Km kcal Z-Gly-Tyr-Phe-Gly-OH 35¡É 2.2 5.82 ¡¿ 10-4M 2.06 sec-1 Z-Gly-Tyr-PheGly-OEt 35¡É 2.4 4.14 ¡¿ 10-4M 1.69 sec-1 The data in the table indicate that the susceptibility of synthetic peptides to peptic hydrolysis are relatively large and the change of the carboxyl-terminal group of synthetic substrate from glycine ethyl ester to glycine causes a small decrease in the susceptibility of the L-tyrosyl-L-phenylalanyl bond.

[学科分类] 化学（综合）