Detecting of fibrino(geno)lytic activity of plant latex proteases by a specific fluorogenic substrate and fibrin(ogen)-zymography

Abstract

Plant latex is repeatedly examined in terms of blood clotting and fibrinolytic activity, due to the proteases often contained. Some examples are the latex of Synadenium grantii Hook (Euphorbiaceae) [1], Pergularia extensa (Jacq.) N.E. Br. (Apocynaceae) [2] or Artocarpus heterophyllus Lam. (Moraceae) [3]. Previously we described the in vitro influence of a protease from the latex of Euphorbia mauritanica L. (Euphorbiaceae) on some steps of the coagulation and fibrinolysis process, by using different methods such as a fibrin plate assay, turbidimetry or in-gel-digestion [4]. We examined over hundred species of the family Euphorbiaceae for fibrinolytic activity with the specific fluorogenic substrate Boc-Val-Leu-Lys-AMC. We have combined the results for the most active plant latices with fibrin(ogen) zymography to confirm the results of the screening and to get information whether the active proteases are the same in the different latices. We also wanted to confirm the results of the above methods for the mentioned isolated protease. The zymograms confirm the activity for all latices tested. However, different proteases seem to be responsible for the activity in each latex.