A new method for determination of elastolytic activity using (14C) labeled elastin and its application to leukocytic elastase.

A new, highly sensitive and specific assay for elastolytic activity is described which employs insoluble elastin randomly labeled with [¹⁴C]. The substrate was prepared by labeling amino groups of the protein in vitro with [¹⁴C] methyl groups by reductive alkylation. The substrate was used to quantitate elastolytic activity from human leukocytes and to compare leukocytic elastase with pancreatic elastase. Purified human leukocytic elastase was approximately one-fourth as active as pancreatic elastase. Similar difference between leukocytic elastase and pancreatic elastase activities was found when the enzymes were tested against succinyl-L-alanyl-L-alanyl-L-alanine-p-nitroanilide, but not when t-BOC-L-alanine-p-nitrophenyl ester was used.