Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom
In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein cont...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2013-12-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/5/12/2403 |
| _version_ | 1811279156143456256 |
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| author | Francis Barbosa Ferreira Mário Sérgio Rocha Gomes Dayane Lorena Naves de Souza Sarah Natalie Cirilo Gimenes Letícia Eulalio Castanheira Márcia Helena Borges Renata Santos Rodrigues Kelly Aparecida Geraldo Yoneyama Maria Inês Homsi Brandeburgo Veridiana M. Rodrigues |
| author_facet | Francis Barbosa Ferreira Mário Sérgio Rocha Gomes Dayane Lorena Naves de Souza Sarah Natalie Cirilo Gimenes Letícia Eulalio Castanheira Márcia Helena Borges Renata Santos Rodrigues Kelly Aparecida Geraldo Yoneyama Maria Inês Homsi Brandeburgo Veridiana M. Rodrigues |
| author_sort | Francis Barbosa Ferreira |
| collection | DOAJ |
| description | In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II) and Ovarian Carcinoma (OVCAR-3), whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast) and Sarcoma 180 (TIB-66). The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects. |
| first_indexed | 2024-04-13T00:49:41Z |
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| id | doaj.art-2d0a712fdb884eec9dc45bc50528b79b |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-13T00:49:41Z |
| publishDate | 2013-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-2d0a712fdb884eec9dc45bc50528b79b2022-12-22T03:09:55ZengMDPI AGToxins2072-66512013-12-015122403241910.3390/toxins5122403toxins5122403Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake VenomFrancis Barbosa Ferreira0Mário Sérgio Rocha Gomes1Dayane Lorena Naves de Souza2Sarah Natalie Cirilo Gimenes3Letícia Eulalio Castanheira4Márcia Helena Borges5Renata Santos Rodrigues6Kelly Aparecida Geraldo Yoneyama7Maria Inês Homsi Brandeburgo8Veridiana M. Rodrigues9Institute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilEzequiel Dias Foundation, FUNED, 30510-010 Belo Horizonte-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilInstitute of Genetics and Biochemistry, Federal University of Uberlândia, UFU, 38400902 Uberlândia-MG, BrazilIn this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II) and Ovarian Carcinoma (OVCAR-3), whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast) and Sarcoma 180 (TIB-66). The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects.http://www.mdpi.com/2072-6651/5/12/2403Bothrops pauloensis phospholipase A2molecular cloning |
| spellingShingle | Francis Barbosa Ferreira Mário Sérgio Rocha Gomes Dayane Lorena Naves de Souza Sarah Natalie Cirilo Gimenes Letícia Eulalio Castanheira Márcia Helena Borges Renata Santos Rodrigues Kelly Aparecida Geraldo Yoneyama Maria Inês Homsi Brandeburgo Veridiana M. Rodrigues Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom Toxins Bothrops pauloensis phospholipase A2 molecular cloning |
| title | Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom |
| title_full | Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom |
| title_fullStr | Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom |
| title_full_unstemmed | Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom |
| title_short | Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom |
| title_sort | molecular cloning and pharmacological properties of an acidic pla2 from bothrops pauloensis snake venom |
| topic | Bothrops pauloensis phospholipase A2 molecular cloning |
| url | http://www.mdpi.com/2072-6651/5/12/2403 |
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