Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina

Denegri, Maria Emília Garcia; Acosta, Ofelia; Huancahuire-Vega, Salomón; Martins-de-Souza, Daniel; Marangoni, Sérgio; Maruñak, Silvana; Teibler, Gladys Pamela; Leiva, Laura; Ponce-Soto, Luis Alberto

doi:10.1016/j.toxicon.2010.02.031

Cited by 44 publications

(25 citation statements)

References 48 publications

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“…The molecular masses obtained by mass spectrometry showed to be similar to that of other snake venom PLA 2 s [22, 24, 25]. The amino acid composition of the Bbil-TX PLA 2 toxin suggests the presence of 14 half-Cys residues, providing the basis for a common structural feature of PLA 2 in the formation of its seven disulfide bridges [20, 21, 26] and a high content of basic and hydrophobic residues, that provides a explication important in the interaction of the PLA 2 with negatively charged phospholipids of cells membranes [27].…”

Section: Discussionmentioning

confidence: 64%

Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX fromBothriopsis bilineataSnake Venom

Carregari

Floriano

Rodrigues‐Simioni

et al. 2013

BioMed Research International

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Bbil-TX, a PLA2, was purified from Bothriopsis bilineata snake venom after only one chromatographic step using RP-HPLC on μ-Bondapak C-18 column. A molecular mass of 14243.8 Da was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode) mass spectrometry. The partial protein sequence obtained was then submitted to BLASTp, with the search restricted to PLA2 from snakes and shows high identity values when compared to other PLA2s. PLA2 activity was presented in the presence of a synthetic substrate and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0 and 25–37°C. Maximum PLA2 activity required Ca2+ and in the presence of Cd2+, Zn2+, Mn2+, and Mg2+ it was reduced in the presence or absence of Ca2+. Crotapotin from Crotalus durissus cascavella rattlesnake venom and antihemorrhagic factor DA2-II from Didelphis albiventris opossum sera under optimal conditions significantly inhibit the enzymatic activity. Bbil-TX induces myonecrosis in mice. The fraction does not show a significant cytotoxic activity in myotubes and myoblasts (C2C12). The inflammatory events induced in the serum of mice by Bbil-TX isolated from Bothriopsis bilineata snake venom were investigated. An increase in vascular permeability and in the levels of TNF-a, IL-6, and IL-1 was was induced. Since Bbil-TX exerts a stronger proinflammatory effect, the phospholipid hydrolysis may be relevant for these phenomena.

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Section: Discussionmentioning

confidence: 64%

Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX fromBothriopsis bilineataSnake Venom

Carregari

Floriano

Rodrigues‐Simioni

et al. 2013

BioMed Research International

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“…Other PLA 2 s have also been purified using simplified procedures based in CM-Sepharose and/or reverse phase, as for example, venom PLA 2 s from Bothrops moojeni [21, 32, 33], B. pirajai [34, 35], B. jararacussu [36, 37], B. alternatus [8, 38], Cerastes cerastes [39], and Elaphe climacophora [40]. …”

Section: Resultsmentioning

confidence: 99%

Purification and Biochemical Characterization of Three Myotoxins fromBothrops mattogrossensisSnake Venom with Toxicity againstLeishmaniaand Tumor Cells

Moura

Kayano

Oliveira

et al. 2014

BioMed Research International

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Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s), one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like), BmatTX-II (Lys49 PLA2-like), and BmatTX-III (Asp49 PLA2). The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T) and SK-BR-3 (breast adenocarcinoma) cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.

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“…However, PLA 2 activity was reduced when BpPLA 2 -TXI was subjected to 100 °C for 30 min, suggesting that denaturation causes loss of protein stability, thereby decreasing its activity. The majority of PLA 2 s from snake venoms are highly stable due to the presence of intra-chain disulfide bonds present in their structures [8,13]. …”

Section: Resultsmentioning

confidence: 99%

Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom

Ferreira

Gomes

Souza

et al. 2013

Toxins

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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.

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Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina

Cited by 44 publications

References 48 publications

Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX fromBothriopsis bilineataSnake Venom

Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX fromBothriopsis bilineataSnake Venom

Purification and Biochemical Characterization of Three Myotoxins fromBothrops mattogrossensisSnake Venom with Toxicity againstLeishmaniaand Tumor Cells

Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom

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