Insulin-like Activity of Proteases. VI. Purification and Some Properties of an Acid-stable N-Succinyl-trialanine p-Nitroanilide-hydrolyzing Protease from Pronase

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An acid-stable N-succinyl-L-trialanine p-nitroanilide-hydrolyzing protease was purified from Pronase E with a protein yield of 6% by affinity chromatography on arginine peptides-Sepharose or soybean trypsin inhibitor-Sepharose. The enzyme showed a potent increasing effect on the glycogen content in an isolated mouse diaphragm. It was homogeneous both in disc and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, had an optimum pH of 8.5,and was stable in the pH range from 2 to 10,whereas its activity was lost at pH 12. Its molecular weight was estimated to be 20800. The sequence up to thirty-two residues from the N-terminus was determined with guanidine-treated and S-carboxymethylated enzymes. The N-and C-terminal residues were isoleucine and tyrosine, respectively. The results suggest that this enzyme is mainly responsible for the insulin-like activity of Pronase and that it is homologous with Protease B which was purified from Pronase B (L. Jurasek, M. R. Carpenter, L. B. Smille, A. Gertler, S. Levy, and L. H. Ericsson, Biochem. Biophys. Res. Commun., 61,1095 (1974)), though some differences were found between the C-terminal regions of the two enzymes.
公益社団法人日本薬学会の論文
1982-11-25