Snake Venom L-Amino Acid Oxidases: Some Consideration About their Functional Characterization

Snake Venom L-amino acid oxidases (LAAOs E.C. 1.4.3.2) are flavoenzymes broadly found in various snake venom compositions. LAAOs have become an attractive subject for molecular biology, biochemistry, physiology and medicine due to their actions on various cells and biological effects on platelets, apoptosis, hemorrhage and others. In this review we try to summarize some of these reports, with special emphasis on apoptosis, anti-protozoa, bactericidal and anti-viral activities.CNPqFINEPFAPESP, Brazi

ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

Submitted by EMERSON LEAL ([email protected]) on 2016-07-05T14:51:42Z No. of bitstreams: 1 ESI-MS MS Identification of a Bradykinin.pdf: 322142 bytes, checksum: 5b32610ed4596f572c944fea47014170 (MD5)Approved for entry into archive by EMERSON LEAL ([email protected]) on 2016-07-14T15:26:16Z (GMT) No. of bitstreams: 1 ESI-MS MS Identification of a Bradykinin.pdf: 322142 bytes, checksum: 5b32610ed4596f572c944fea47014170 (MD5)Made available in DSpace on 2016-07-14T15:26:16Z (GMT). No. of bitstreams: 1 ESI-MS MS Identification of a Bradykinin.pdf: 322142 bytes, checksum: 5b32610ed4596f572c944fea47014170 (MD5) Previous issue date: 2013Made available in DSpace on 2016-07-15T18:40:38Z (GMT). No. of bitstreams: 3 ESI-MS MS Identification of a Bradykinin.pdf.txt: 21633 bytes, checksum: 54e00469ca67e58fb4b3385fb3bd3091 (MD5) ESI-MS MS Identification of a Bradykinin.pdf: 322142 bytes, checksum: 5b32610ed4596f572c944fea47014170 (MD5) license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5) Previous issue date: 2013Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Waters Corporation. MS Applications Development Laboratory. Alphaville, SP, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil / Federal University of Rondônia. Medicine Department. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil / Federal University of Rondônia. Medicine Department. Porto Velho, RO, Brasil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Center of Biomolecules Study Applied to Health. Porto Velho, RO, Brasil. / Federal University of Rondônia. Medicine Department. Porto Velho, RO, Brasil.A bradykinin-potentiating peptide (BPP) from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID) using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK). De novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is similar to experimental data, showing a mass accuracy of 0.6 ppm. The peptide is homologous to other BPP from Bothrops moojeni and was named as BPP-BAX12

Activation of J77A.1 macrophages by three phospholipases A2 isolated from bothrops atrox snake venom

Submitted by Claudete Queiroz ([email protected]) on 2016-05-03T17:00:14Z No. of bitstreams: 1 Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom.pdf: 2090853 bytes, checksum: 0669d064c65a39c61c661542c067ee9b (MD5)Approved for entry into archive by EMERSON LEAL ([email protected]) on 2016-05-10T14:32:57Z (GMT) No. of bitstreams: 1 Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom.pdf: 2090853 bytes, checksum: 0669d064c65a39c61c661542c067ee9b (MD5)Made available in DSpace on 2016-05-10T14:32:57Z (GMT). No. of bitstreams: 1 Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom.pdf: 2090853 bytes, checksum: 0669d064c65a39c61c661542c067ee9b (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil. / Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Laboratório de Imunofarmacologia Aplicada à Saúde. Porto Velho, RO, Brazil. / Fundação Oswaldo Cruz. Centro de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.In the present study, we investigated the in vitro effects of two basic myotoxic phospholipases A2 (PLA2), BaTX-I, a catalytically inactive Lys-49 variant, and BaTX-II, a catalytically active Asp-49, and of one acidic myotoxic PLA2, BaPLA2, a catalytically active Asp-49, isolated from Bothrops atrox snake venom, on the activation of J774A.1 macrophages. At noncytotoxic concentrations, the toxins did not affect the adhesion of the macrophages, nor their ability to detach. The data obtained showed that only BaTXI stimulated complement receptor-mediated phagocytosis. However, BaTX-I, BaTX-II, and BaPLA2 induced the release of the superoxide anion by J774A.1 macrophages. Additionally, only BaTX-I raised the lysosomal volume of macrophages after 15 min of incubation. After 30 min, all the phospholipases increased this parameter, which was not observed within 60 min. Moreover, BaTX-I, BaTX-II, and BaPLA2 increased the number of lipid bodies on macrophages submitted to phagocytosis and not submitted to phagocytosis. However, BaTX-II and BaPLA2 induced the release of TNF- by J774A.1 macrophages. Taken together, the data show that, despite differences in enzymatic activity, the three toxins induced inflammatory events and whether the enzyme is acidic or basic does not seem to contribute to these effects

Antitumoral Activity of Snake Venom Proteins: New Trends in Cancer Therapy

For more than half a century, cytotoxic agents have been investigated as a possible treatment for cancer. Research on animal venoms has revealed their high toxicity on tissues and cell cultures, both normal and tumoral. Snake venoms show the highest cytotoxic potential, since ophidian accidents cause a large amount of tissue damage, suggesting a promising utilization of these venoms or their components as antitumoral agents. Over the last few years, we have studied the effects of snake venoms and their isolated enzymes on tumor cell cultures. Some in vivo assays showed antineoplastic activity against induced tumors in mice. In human beings, both the crude venom and isolated enzymes revealed antitumor activities in preliminary assays, with measurable clinical responses in the advanced treatment phase. These enzymes include metalloproteases (MP), disintegrins, L-amino acid oxidases (LAAOs), C-type lectins, and phospholipases A2 (PLA2s). Their mechanisms of action include direct toxic action (PLA2s), free radical generation (LAAOs), apoptosis induction (PLA2s, MP, and LAAOs), and antiangiogenesis (disintegrins and lectins). Higher cytotoxic and cytostatic activities upon tumor cells than normal cells suggest the possibility for clinical applications. Further studies should be conducted to ensure the efficacy and safety of different snake venom compounds for cancer drug development

Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom

Submitted by Claudete Queiroz ([email protected]) on 2016-05-02T01:48:25Z No. of bitstreams: 1 Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom.pdf: 4262246 bytes, checksum: d6e3f5bb80b013a337bed40150e217dd (MD5)Approved for entry into archive by EMERSON LEAL ([email protected]) on 2016-05-03T13:19:15Z (GMT) No. of bitstreams: 1 Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom.pdf: 4262246 bytes, checksum: d6e3f5bb80b013a337bed40150e217dd (MD5)Made available in DSpace on 2016-05-03T13:19:15Z (GMT). No. of bitstreams: 1 Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom.pdf: 4262246 bytes, checksum: d6e3f5bb80b013a337bed40150e217dd (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Universidade Federal Fluminense. Instituto de Biologia. Departamento de Biologia Celular e Molecular. Niteroi, RJ, Brazil.Universidade Federal do ABC. Centro de Ciências Naturais e Humanas. Santo André, SP, Brazil.Universidade Federal do ABC. Centro de Ciências Naturais e Humanas. Santo André, SP, Brazil. / Universidade de São Paulo. Escola de Artes, Ciências e Humanidades. São Paulo, SP, Brazil.Universidade Federal de São João Del Rei. Campus Alto Paraopeba. Ouro Branco, MG, Brazil.Institute for Research in Biomedicine. Barcelona, Spain. / Barcelona Science Park. Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina. Barcelona, Spain.Barcelona Science Park. Proteomic Platform. Barcelona, Spain.Barcelona Science Park. Proteomic Platform. Barcelona, Spain.Institute for Research in Biomedicine. Barcelona, Spain. / Barcelona Science Park. Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina. Barcelona, Spain. / University of Barcelona. Department of Organic Chemistry. Barcelona, Spain. / University of KwaZulu Natal. School of Chemistry. Durban, South Africa.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde. Departamento de Medicina. Universidade Federal de Rondônia. Porto Velho, RO, Brazil.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 (39,408.32Da). 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, un like most serine proteases. BpirSP-39 was capable of hydrolyzing different chromogenic substrates tested (S-2222, S-2302, and S-2238) while Cu2+ 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