Isolation and biochemical characterization of a new thrombin-like serine protease from Bothrops pirajai snake venom.
This paper presents a novel serine protease (SP) isolated from Bothrops pirajai, a venomous snake found solely in Brazil that belongs to the Viperidae family. The identified SP, named BpirSP-39, was isolated by three chromatographic steps (size exclusion, bioaffinity, and reverse phase chromatographies). The molecular mass of BpirSP-39 was estimated by SDS-PAGE and confirmed by mass spectrometry (39,408.32 Da). The protein was able to form fibrin networks, which was not observed in the presence of serine protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF). Furthermore, BpirSP-39 presented considerable thermal stability and was apparently able to activate factor XIII of the blood coagulation cascade, unlike most serine proteases. BpirSP-39 was capable of hydrolyzing different chromogenic substrates tested (S-2222, S-2302, and S-2238) while Cu(2+) significantly diminished BspirSP-39 activity on the three tested substrates. The enzyme promoted platelet aggregation and also exhibited fibrinogenolytic, fibrinolytic, gelatinolytic, and amidolytic activities. The multiple alignment showed high sequence similarity to other thrombin-like enzymes from snake venoms. These results allow us to conclude that a new SP was isolated from Bothrops pirajai snake venom.
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Kayena D
KD
Zaqueo
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Anderson M
AM
Kayano
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Rodrigo
R
Simões-Silva
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Leandro S
LS
Moreira-Dill
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Carla F C
CF
Fernandes
Departmento de Biologia Celular e Molecular, Instituto de Biologia, Universidade Federal Fluminense, 24210-130 Niteroi, RJ, Brazil.
André L
AL
Fuly
Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, 09210-170 Santo André, SP, Brazil.
Vinícius G
VG
Maltarollo
Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, 09210-170 Santo André, SP, Brazil ; Escola de Artes, Ciências e Humanidades, USP, 03828-000 São Paulo, SP, Brazil.
Kathia M
KM
Honório
Universidade Federal de São João Del Rei, UFSJ, Campus Alto Paraopeba, 36420-000 Ouro Branco, MG, Brazil.
Saulo L
SL
da Silva
Institute for Research in Biomedicine (IRB Barcelona), 08028 Barcelona, Spain ; CIBER-BBN, Barcelona Science Park, 08028 Barcelona, Spain.
Gerardo
G
Acosta
Proteomic Platform, Barcelona Science Park, 08028 Barcelona, Spain.
Maria Antonia O
MA
Caballol
Proteomic Platform, Barcelona Science Park, 08028 Barcelona, Spain.
Eliandre
E
de Oliveira
Institute for Research in Biomedicine (IRB Barcelona), 08028 Barcelona, Spain ; CIBER-BBN, Barcelona Science Park, 08028 Barcelona, Spain ; Department of Organic Chemistry, University of Barcelona, 08028 Barcelona, Spain ; School of Chemistry, University of KwaZulu Natal, Durban 4001, South Africa.
Fernando
F
Albericio
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Leonardo A
LA
Calderon
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Andreimar M
AM
Soares
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, Fiocruz Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, UNIR, Rua da Beira 7176, Bairro Lagoa, 76812-245 Porto Velho, RO, Brazil.
Rodrigo G
RG
Stábeli
D000818:Animals; D017837:Bothrops; D003435:Crotalid Venoms; D005169:Factor VIII / Q000737:chemistry*; D005342:Fibrinolysis; D006801:Humans; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000737:chemistry; D057057:Serine Proteases / Q000737:chemistry* / Q000302:isolation & purification*; D015842:Serine Proteinase Inhibitors / Q000737:chemistry
D003435:Crotalid Venoms; D015842:Serine Proteinase Inhibitors; D010664:Phenylmethylsulfonyl Fluoride; C078147:F8 protein, human; D005169:Factor VIII; D057057:Serine Proteases