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Comp Biochem Physiol A Mol Integr Physiol. 2016 May;195:15-25. doi: 10.1016/j.cbpa.2016.01.021. Epub 2016 Jan 28.

BbrzSP-32, the first serine protease isolated from Bothrops brazili venom: Purification and characterization.

Author information

1
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
2
Universidade Federal Fluminense, UFF, Niterói, RJ, Brazil.
3
Universidade Regional Amazónica IKIAM, Via Muyuna Km 7, Tena, Napo, Ecuador.
4
Institute for Research in Biomedicine (IRB Barcelona), 08028-Barcelona, Spain and CIBER-BBN, Barcelona Science Park, Barcelona 08028, Spain.
5
Proteomic Platform, Barcelona Science Park, Barcelona 08028, Spain.
6
Institute for Research in Biomedicine (IRB Barcelona), 08028-Barcelona, Spain and CIBER-BBN, Barcelona Science Park, Barcelona 08028, Spain; Department of Organic Chemistry University of Barcelona, Barcelona 08028, Spain; School of Chemistry, University of KwaZulu Natal, Durban 4001, South Africa.
7
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil. Electronic address: stabeli@fiocruz.br.
8
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil. Electronic address: andreimar@fiocruz.br.

Abstract

Snake venom toxins are related not only in detention, death and the promotion of initial digestion of prey but also due to their different biochemical, structural and pharmacological effects they can result in new drugs. Among these toxins snake venom serine proteases (SVSPs) should be highlighted because they are responsible for inducing changes in physiological functions such as blood coagulation, fibrinolysis, and platelet aggregation. This article presents the first serine protease (SP) isolated from Bothrops brazili: BbrzSP-32. The new SP showed 36 kDa of relative molecular mass and its absolute mass was confirmed by mass spectrometry as 32,520 Da. It presents 79.48% identity when compared to other SVSPs and was able to degrade the α-chain of fibrinogen, in in vitro models, because of this it is considered a SVTLE-A. It showed dose-dependent activity in the process of degradation of fibrin networks demonstrating greater specificity for this activity when compared to its thrombolytic action. BbrzSP-32 demonstrated proteolytic activity on gelatin and chromogenic substrates for serine proteases and thrombin-like enzymes (S-2288 and S-2238 respectively), besides having coagulant activity on human plasma. After pre-incubation with PMSF and benzamidine the coagulant and proteolytic activities on the S-2288 and S-2238 substrates were reduced. BbrzSP-32 shows stability against pH and temperature variations, demonstrating optimum activity between 30 and 40 °C and in the pH range 7.5 to 8.5. A new SP with potential biotechnological application was isolated.

KEYWORDS:

Biotechnology; Enzyme; Snake venom; Thrombin-like; Viperidae

PMID:
26827743
DOI:
10.1016/j.cbpa.2016.01.021
[Indexed for MEDLINE]
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