Isolation of a myotoxin from Bothrops asper venom: Partial characterization and action on skeletal muscle

Gutiérrez, José Marı́a; Ownby, Charlotte L.; Odell, George V.

doi:10.1016/0041-0101(84)90144-2

Cited by 155 publications

(73 citation statements)

References 30 publications

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“…Attempts using amino-acid sequence comparisons to identify the region of the PLA2 responsible for myotoxicity (Kini & Iwanaga,986) have not been successful in predicting myotoxity in myotoxin II (Francis et al, 1991). The same degree of myotoxicity and edema-inducing activity exists between myotoxins I and II (Gutierrez, Ownby & Odell, 1984), suggesting that phospholipase activity is not essential for these functions. Previous comparisons of myotoxic PLA2 amino-acid sequences indicate that all have either three or four tyrosines between residues 112 and 121, a threonine at 112, and a lysine at position 38, all of which are not found in non-myotoxic PLA2 sequences (Francis et al, 1991).…”

Section: S48mentioning

confidence: 99%

Structure of a calcium-independent phospholipase-like myotoxic protein fromBothrops aspervenom

Arni¹,

Ward²,

Gutiérrez³

et al. 1995

Acta Crystallogr D Biol Crystallogr

104

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Myotoxin II, a myotoxic calcium-independent phospholipase-like protein isolated from the venom of Bothrops asper, possesses no detectable phospholipase activity. The crystal structure has been determined and refined at 2.8A to an R factor of 16.5% (F>3o') with excellent stereochemistry. Amino-acid differences between catalytically active phospholipases and myotoxin II in the Ca2+-binding region, specifically the substitutions Tyr28~Asn, Gly32~Leu and Asp49~Lys, result in an altered local conformation. The key difference is that the e-amino group of Lys49 fills the site normally occupied by the calcium ion in catalytically active phospholipases. In contrast to the homologous monomeric Lys49 variant from Agkistrodon piscivorus piscivorus, myotoxin II is present as a dimer both in solution and in the crystalline state. The two molecules in the asymmetric unit are related by a nearly perfect twofold axis, yet the dimer is radically different from the dimer formed by the phospholipase from Crotalus atrox. Whereas in C. atrox the dimer interface occludes the active sites, in myotoxin II they are exposed to solvent.

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Section: S48mentioning

confidence: 99%

Structure of a calcium-independent phospholipase-like myotoxic protein fromBothrops aspervenom

Arni¹,

Ward²,

Gutiérrez³

et al. 1995

Acta Crystallogr D Biol Crystallogr

104

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show abstract

“…The unbound fraction was collected, freeze-dried, and stored at À20 C. Homogeneity of the final preparation was evaluated by sodium dodecylsulphate-polyacrylamide (15%) gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions, followed by Coomassie blue R-250 staining. In some experiments, two basic PLA 2 s from B. asper venom were included for comparative purposes: myotoxin I is a catalytically active Asp49 enzyme [27], whereas myotoxin II is a catalytically inactive Lys49 homologue [28].…”

Section: Isolation Of Basppla 2 -Iimentioning

confidence: 99%

Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: Biochemical and toxicological characterization☆

et al. 2010

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a b s t r a c tPhospholipases A 2 (PLA 2 ) are major components of snake venoms, exerting a variety of relevant toxic actions such as neurotoxicity and myotoxicity, among others. Since the majority of toxic PLA 2 s are basic proteins, acidic isoforms and their possible roles in venoms are less understood. In this study, an acidic enzyme (BaspPLA 2 -II) was isolated from the venom of Bothrops asper (Pacific region of Costa Rica) and characterized. BaspPLA 2 -II is monomeric, with a mass of 14,212 AE 6 Da and a pI of 4.9. Its complete sequence of 124 amino acids was deduced through cDNA and protein sequencing, showing that it belongs to the Asp49 group of catalytically active enzymes. In vivo and in vitro assays demonstrated that BaspPLA 2 -II, in contrast to the basic Asp49 counterparts present in the same venom, lacks myotoxic, cytotoxic, and anticoagulant activities. BaspPLA 2 -II also differed from other acidic PLA 2 s described in Bothrops spp. venoms, as it did not show hypotensive and anti-platelet aggregation activities. Furthermore, this enzyme was not lethal to mice at intravenous doses up to 100 mg (5.9 mg/g), indicating its lack of neurotoxic activity. The only toxic effect recorded in vivo was a moderate induction of local edema. Therefore, the toxicological characteristics of BaspPLA 2 -II suggest that it does not play a key role in the pathophysiology of envenomings by B. asper, and that its purpose might be restricted to digestive functions. Immunochemical analyses using antibodies raised against BaspPLA 2 -II revealed that acidic and basic PLA 2 s form two different antigenic groups in B. asper venom.

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“…Así por ejemplo, podemos mencionar las miotoxinas aisladas de los venenos de B. asper, B. nummifer y B. moojeni, (Gutierrez et al, 1984;Gutierrez et al, 1986;Lomonte et al, 1990;Soares et al, 1998).…”

Section: Resultados Y Discusión 1 Purificación De La Miotoxinaunclassified

“…3), lo cual representa un tiempo similar al encontrado con la miotoxina I de B. asper y la miotoxina II de B. mojeni, las cuales provocan la liberación máxima de creatina kinasa recién a las tres horas (Gutierrez et al, 1984;Lomonte et al, 1990). Sin embargo, los niveles de creatina kinasa prácticamente se normalizan luego de 24 horas.…”

Section: Actividad Miotóxicaunclassified

Aislamiento y caracterización parcial de una miotoxina del veneno de la serpiente Bothrops atrox (Ophidia: Viperidae)

Huatuco

Escobar²,

Yarlequé

2004

Rev peru biol

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ResumenSe ha purificado una miotoxina del veneno de la serpiente Bothrops atrox, empleando una columna de intercambio catiónico de CM-Sephadex C-50 equilibrada con buffer acetato de amonio 0,05M a pH 7. La miotoxina es una proteína básica, y por cromatografía de filtración y PAGE-SDS se ha determinado que tiene un peso molecular de 27 kDa, estando formada por dos cadenas polipeptídicas de 14 kDa cada una.La inoculación de la miotoxina en el músculo gastrocnemius de ratones albinos produce la liberación de creatina kinasa así como la necrosis del tejido. La miotoxina tiene actividad de fosfolipasa, anticoagulante y edemática, pero no hemolítica. Palabras claves: miotoxina, Bothrops atrox, veneno de serpiente, mionecrosis AbstractA myotoxin from Bothrops atrox snake venom was purified by cationic exchange on CM-Sephadex C-50 with 0,05M ammonium acetate pH 7. The myotoxin is a basic protein and by gel filtration and PAGE-SDS was demonstrated that protein has a molecular weight of 27 kDa and two polipeptides chain of 14 kDa each one.The inoculation of myotoxin in gastrocnemius muscle of white mice produce liberation of creatin kinase as well as myonecrosis. The myotoxin has phospholipasic, anticoagulant and edematic activity, but not hemolytic activity.

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Isolation of a myotoxin from Bothrops asper venom: Partial characterization and action on skeletal muscle

Cited by 155 publications

References 30 publications

Structure of a calcium-independent phospholipase-like myotoxic protein fromBothrops aspervenom

Structure of a calcium-independent phospholipase-like myotoxic protein fromBothrops aspervenom

Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: Biochemical and toxicological characterization☆

Aislamiento y caracterización parcial de una miotoxina del veneno de la serpiente Bothrops atrox (Ophidia: Viperidae)

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