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

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

BIOCHIMIE

ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

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

0300-9084

Plant serine proteases have been proposed as potential anti-coagulants and anti-platelet agents. Recently, we have reported the purification and identification of a Solanum tuberosum subtilisin-like protease, named as StSBTc-3. Here we report that, like several plant serine proteases, this enzyme exerts fibrino(geno)lytic and anti-platelet activities. Results obtained show that StSBTc-3 degrades all chains of fibrinogen and to produce fibrin clot lysis in a dose dependent manner, at similar amounts to those previously reported for other plant serine proteases. Evaluation of biochemical properties of StSBTc-3 fibrinolytic activity determinate that, the optimal pH is 8 and the optimal temperature is 37 C. Fibrinogenolytic activity of StSBTc- 3 is stable between pH 5.0 to 12.0 and at temperature from 30 C to 60 C. Determination of StSBTc- 3 substrate specificity on oxidized insulin â chain shows seven cleavage sites (Asn3-Gln4; Cys7-Gly8; Glu13-Ala14; Leu15-Tyr16; Tyr16-Leu17; Arg22-Gly23 and Phe25-Tyr26), all of them were previously described to other serine proteases with fibrinogenolytic. These results could be explaining the substrate specificity of StSBTc- 3 on fibrinogen chains. Additionally, data obtained in this work show that StSBTc-3 inhibits platelet aggregation and is unable to exert cytotoxic activity on human erythrocytes in vitro. All 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.