Differentiation of the elastase-type protease of platelets from other elastases.

Elastase activities were determined near neutral pH on several specific substrates using platelet-derived preparations mixed with decreasing amounts of leukocytes. Activities were extrapolated to zero leukocyte content enabling the estimation of intrinsic platelet elastase activity. In contrast to human leukocyte elastase, metal chelating agents inhibited partly the elastase activity of the platelet extract and soybean trypsin inhibitor did not modify its activity. Serine active site titrants (phenylmethane sulfonyl fluoride) as well as acetyl-di-L-alanyl-L-propyl-L-valine chloromethylketone completely abolished the activity of platelet lysates. The platelet protease was purified from Triton X-100 platelet lysates. No cross-reactivity could be demonstrated by immunoelectrophoresis with either porcine pancreatic elastase or human leukocyte elastase using monospecific antisera. Applying gel electrophoresis, most of the elastase activity of the platelet protease migrated towards the anode, whereas the pancreatic and leukocyte elastases migrated towards the cathode. The anionic character of the platelet enzyme might explain its capacity to degrade better elastin treated with cationic detergents in contradistinction to other elastases which act better on anionic detergent-treated elastins.