title: Cotiarinase is a novel prothrombin activator from the venom of Bothrops cotiara.
abstract: Snake venom serine proteinases (SVSPs) may affect hemostatic pathways by specifically activating components involved in coagulation, fibrinolysis and platelet aggregation or by unspecific proteolytic degradation. In this study, we purified and characterized an SVSP from Bothrops cotiara venom, named cotiarinase, which generated thrombin upon incubation with prothrombin. Cotiarinase was isolated by a two-step procedure including gel-filtration and cation-exchange chromatographies and showed a single protein band with a molecular mass of 29 kDa by SDS-polyacrylamide gel electrophoresis under reducing conditions. Identification of cotiarinase by mass spectrometric analysis revealed peptides that matched sequences of viperid SVSPs. Cotiarinase did not show fibrinogen-clotting, platelet-aggregating, fibrinogenolytic and factor X activating activities. Upon incubation with prothrombin the generation of thrombin was detected using the peptide substrate d-Phe-Pip-Arg-pNA. Moreover, mass spectrometric identification of prothrombin fragments generated by cotiarinase in the absence of co-factors (phospholipids, factor Va, factor Xa and Ca(2+) ions), indicated the limited proteolysis of this protein to release prothrombin 1, fragment 1 and thrombin. Cotiarinase is a novel SVSP that acts on prothrombin to release active thrombin that does not match any group of the current classification of snake venom prothrombin activators.
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, CAT-cepid, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, Brazil.
authors.forename: Eduardo S
authors.initials: ES
authors.lastname: Kitano
authors.forename: Thalita C
authors.initials: TC
authors.lastname: Garcia
authors.forename: Milene C
authors.initials: MC
authors.lastname: Menezes
authors.forename: Alexandre K
authors.initials: AK
authors.lastname: Tashima
authors.forename: André
authors.initials: A
authors.lastname: Zelanis
authors.forename: Solange M T
authors.initials: SM
authors.lastname: Serrano
mesh_terms: D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D011516:Prothrombin / Q000378:metabolism*; D057057:Serine Proteases / Q000302:isolation & purification / Q000378:metabolism*; D012910:Snake Venoms / Q000201:enzymology*
chemical_list: D012910:Snake Venoms; D011516:Prothrombin; D057057:Serine Proteases