title: Moojenactivase, a novel pro-coagulant PIIId metalloprotease isolated from Bothrops moojeni snake venom, activates coagulation factors II and X and induces tissue factor up-regulation in leukocytes.
abstract: Coagulopathies following snakebite are triggered by pro-coagulant venom toxins, in which metalloproteases play a major role in envenomation-induced coagulation disorders by acting on coagulation cascade, platelet function and fibrinolysis. Considering this relevance, here we describe the isolation and biochemical characterization of moojenactivase (MooA), a metalloprotease from Bothrops moojeni snake venom, and investigate its involvement in hemostasis in vitro. MooA is a glycoprotein of 85,746.22 Da, member of the PIIId group of snake venom metalloproteases, composed of three linked disulfide-bonded chains: an N-glycosylated heavy chain, and two light chains. The venom protease induced human plasma clotting in vitro by activating on both blood coagulation factors II (prothrombin) and X, which in turn generated α-thrombin and factor Xa, respectively. Additionally, MooA induced expression of tissue factor (TF) on the membrane surface of peripheral blood mononuclear cells (PBMC), which led these cells to adopt pro-coagulant characteristics. MooA was also shown to be involved with production of the inflammatory mediators TNF-α, IL-8 and MCP-1, suggesting an association between MooA pro-inflammatory stimulation of PBMC and TF up-regulation. We also observed aggregation of washed platelets when in presence of MooA; however, the protease had no effect on fibrinolysis. Our findings show that MooA is a novel hemostatically active metalloprotease, which may lead to the development of coagulopathies during B. moojeni envenomation. Moreover, the metalloprotease may contribute to the development of new diagnostic tools and pharmacological approaches applied to hemostatic disorders.
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Marco A
authors.initials: MA
authors.lastname: Sartim
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Tassia R
authors.initials: TR
authors.lastname: Costa
authors.affiliation: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos e Centro de Química de Proteínas, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Helen J
authors.initials: HJ
authors.lastname: Laure
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Milena S
authors.initials: MS
authors.lastname: Espíndola
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Fabiani G
authors.initials: FG
authors.lastname: Frantz
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Sorgi
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Adélia C O
authors.initials: AC
authors.lastname: Cintra
authors.affiliation: Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Eliane C
authors.initials: EC
authors.lastname: Arantes
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: Lucia H
authors.initials: LH
authors.lastname: Faccioli
authors.affiliation: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos e Centro de Química de Proteínas, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil.
authors.forename: José C
authors.initials: JC
authors.lastname: Rosa
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-903, Brazil. suvilela@usp.br.
authors.forename: Suely V
authors.initials: SV
authors.lastname: Sampaio
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D017837:Bothrops / Q000378:metabolism*; D003029:Coagulants / Q000302:isolation & purification / Q000494:pharmacology*; D003435:Crotalid Venoms / Q000201:enzymology* / Q000302:isolation & purification / Q000494:pharmacology; D004795:Enzyme Stability; D015951:Factor Xa / Q000378:metabolism*; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018836:Inflammation Mediators / Q000378:metabolism; D007700:Kinetics; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D045726:Metalloproteases / Q000302:isolation & purification / Q000494:pharmacology*; D008875:Middle Aged; D011516:Prothrombin / Q000378:metabolism*; D013696:Temperature; D013925:Thromboplastin / Q000378:metabolism*; D055815:Young Adult
chemical_list: D003029:Coagulants; D003435:Crotalid Venoms; D018836:Inflammation Mediators; D011516:Prothrombin; C025337:Factor IIa; D013925:Thromboplastin; D045726:Metalloproteases; D015951:Factor Xa; D008666:Metalloendopeptidases; C000613573:moojenactivase, Bothrops moojeni
keywords: Bothrops moojeni; Coagulation factors; Metalloprotease; Platelet aggregation; Pro-coagulant; Tissue factor