Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom
A new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homolo...
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | Biochemistry Research International |
| Online Access: | http://dx.doi.org/10.1155/2015/826059 |
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| author | Georgina Sucasaca-Monzón Priscila Randazzo-Moura Thalita Rocha Frank Denis Torres-Huaco Augusto Vilca-Quispe Luis Alberto Ponce-Soto Sérgio Marangoni Maria Alice da Cruz-Höfling Léa Rodrigues-Simioni |
| author_facet | Georgina Sucasaca-Monzón Priscila Randazzo-Moura Thalita Rocha Frank Denis Torres-Huaco Augusto Vilca-Quispe Luis Alberto Ponce-Soto Sérgio Marangoni Maria Alice da Cruz-Höfling Léa Rodrigues-Simioni |
| author_sort | Georgina Sucasaca-Monzón |
| collection | DOAJ |
| description | A new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 PLA2 myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°–45°C. Full Bp-13 PLA2 activity required Ca2+; its PLA2 activity was inhibited by Mg2+, Mn2+, Sr2+, and Cd2+ in the presence and absence of 1 mM Ca2+. In the mouse phrenic nerve-diaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (P<0.05). Both the replacement of Ca2+ by Sr2+ and temperature lowering (24°C) inhibited the Bp-13 PLA2-induced twitch-tension blockade. Bp-13 PLA2 inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (P>0.05). The main effect of this new Asp49 PLA2 of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom. |
| format | Article |
| id | doaj-art-d08ef520bc2b43009ea15656a436990b |
| institution | Kabale University |
| issn | 2090-2247 2090-2255 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Biochemistry Research International |
| spelling | doaj-art-d08ef520bc2b43009ea15656a436990b2025-02-03T01:24:23ZengWileyBiochemistry Research International2090-22472090-22552015-01-01201510.1155/2015/826059826059Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake VenomGeorgina Sucasaca-Monzón0Priscila Randazzo-Moura1Thalita Rocha2Frank Denis Torres-Huaco3Augusto Vilca-Quispe4Luis Alberto Ponce-Soto5Sérgio Marangoni6Maria Alice da Cruz-Höfling7Léa Rodrigues-Simioni8Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), 13083-881 Campinas, SP, BrazilDepartment of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), 13083-881 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Biochemistry and Tissue Biology, Institute of Biology, State University of Campinas (UNICAMP), 13083-365 Campinas, SP, BrazilDepartment of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), 13083-881 Campinas, SP, BrazilA new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 PLA2 myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°–45°C. Full Bp-13 PLA2 activity required Ca2+; its PLA2 activity was inhibited by Mg2+, Mn2+, Sr2+, and Cd2+ in the presence and absence of 1 mM Ca2+. In the mouse phrenic nerve-diaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (P<0.05). Both the replacement of Ca2+ by Sr2+ and temperature lowering (24°C) inhibited the Bp-13 PLA2-induced twitch-tension blockade. Bp-13 PLA2 inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (P>0.05). The main effect of this new Asp49 PLA2 of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom.http://dx.doi.org/10.1155/2015/826059 |
| spellingShingle | Georgina Sucasaca-Monzón Priscila Randazzo-Moura Thalita Rocha Frank Denis Torres-Huaco Augusto Vilca-Quispe Luis Alberto Ponce-Soto Sérgio Marangoni Maria Alice da Cruz-Höfling Léa Rodrigues-Simioni Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom Biochemistry Research International |
| title | Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom |
| title_full | Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom |
| title_fullStr | Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom |
| title_full_unstemmed | Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom |
| title_short | Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom |
| title_sort | bp 13 pla2 purification and neuromuscular activity of a new asp49 toxin isolated from bothrops pauloensis snake venom |
| url | http://dx.doi.org/10.1155/2015/826059 |
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