Fibrin(ogen)olytic and antiplatelet activities of a subtilisin-like protease from Solanum tuberosum (StSBTc-3)

ALFONSO PEPE; MARÍA EUGENIA FREY; FERNANDO MUÑOZ; MARÍA BELÉN FERNÁNDEZ; ANABELA PEDRAZA; GUSTAVO GALBÁN; DIANA NOEMÍ GARCÍA; GUSTAVO RAÚL DALEO; MARÍA GABRIELA GUEVARA

BIOCHIMIE

ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

Lugar: Paris; Año: 2016 vol. 125 p. 163 - 170

0300-9084

Plant serine proteases have been widely used in food science and technology as wellas in medicine. In this sense, several plant serine proteases have been proposed as potential anti-coagulants and anti-platelet agents. Previously, we have reported the purification and identication of a plant serine protease fromSolanum tuberosum leaves. This potato enzyme, named as StSBTc-3, has a molecular weight of 72 kDa and it was characterized as a subtilisin likeprotease. In this work we determine and characterize the biochemical and medicinal properties of StSBTc-3. Results obtained show that, like the reportedto other plant serine proteases, StSBTc-3 is able to degrade all chains ofhuman fibrinogen and to produces fibrin clot lysis in a dose dependent manner.The enzyme efficiently hydrolyses beta subunit followed by partially hydrolyzed alpha and gamma subunits of human fibrinogen. Assays performed to determineStSBTc-3 substrate specificity using oxidized insulin beta-chain as substrate,show seven cleavage sites: Asn3-Gln4; Cys7-Gly8; Glu13-Ala14; Leu15-Tyr16;Tyr16- Leu17; Arg22-Gly23 and Phe25-Tyr26, all of them were previously reportedfor other serine proteases with fibrinogenolytic activity. The maximum StSBTc-3fibrinogenolytic activity was determined at pH 8.0 and at 37 C. Additionally, we demonstrate that StSBTc-3 is able to inhibit platelet aggregation and is unable to exert cytotoxic activity on human erythrocytes in vitro at all concentrations assayed. These results suggest that StSBTc-3 could be evaluated as a new agent to be used in the treatment of thromboembolic disorders such as strokes, pulmonary embolism and deep vein thrombosis.