Key events in microvascular damage induced by snake venom hemorrhagic metalloproteinases

Escalante, Teresa; Rucavado, Alexandra; Fox, Jay W.; Gutiérrez, José Marı́a

doi:10.1016/j.jprot.2011.03.026

Cited by 208 publications

(149 citation statements)

References 89 publications

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“…Hemorrhagic P-III SVMPs, which comprise the metalloprotease domain and disintegrin-like (Dis) domain, followed by a cysteine-rich (CR) domain, gather in capillary blood and vessels by binding to the basement membrane through the Dis and CR domains to disrupt the microvascular system very effectively (52). In addition, the noncatalytic Dis and CR domains containing substrate binding sites were considered to confer greater hemorrhagic activity on P-III SVMPs (53). In our study, the mixture of the four scFv antibodies revealed the inhibition of the lethal effect of the DA venom protein in in vivo neutralization assays (Fig.…”

Section: Discussionmentioning

confidence: 99%

Antibodies against Venom of the Snake Deinagkistrodon acutus

Lee

Liang

et al. 2016

Appl Environ Microbiol

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h Snake venom protein from Deinagkistrodon acutus (DA protein), one of the major venomous species in Taiwan, causes hemorrhagic symptoms that can lead to death. Although horse-derived antivenin is a major treatment, relatively strong and detrimental side effects are seen occasionally. In our study, yolk immunoglobulin (IgY) was purified from eggs, and DA protein was recognized using Western blotting and an enzyme-linked immunosorbent assay (ELISA), similar to therapeutic horse antivenin. The ELISA also indicated that specific IgY antibodies were elicited after the fifth booster, plateaued, and lasted for at least 3 months. To generate monoclonal single-chain variable fragment (scFv) antibodies, we used phage display technology to construct two libraries with short or long linkers, containing 6.24 ؋ 10 8 and 5.28 ؋ 10 8 transformants, respectively. After four rounds of biopanning, the eluted phage titer increased, and the phage-based ELISA indicated that the specific clones were enriched. Nucleotide sequences of 30 individual clones expressing scFv were analyzed and classified into four groups that all specifically recognized the DA venom protein. Furthermore, based on mass spectrometry, the scFv-bound protein was deduced to be snake venom metalloproteinase proteins. Most importantly, both IgY and mixed scFv inhibited the lethal effect in mice injected with the minimum lethal dosage of the DA protein. We suggest that together, these antibodies could be applied to the development of diagnostic agents or treatments for snakebite envenomation in the future.

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Section: Discussionmentioning

confidence: 99%

Antibodies against Venom of the Snake Deinagkistrodon acutus

Lee

Liang

et al. 2016

Appl Environ Microbiol

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“…In addition, these venoms contain bradykinin-potentiating peptides (BPPs), disintegrins, C-type lectin-like proteins, L-amino acid oxidase and various other enzymes (Calvete et al, 2009). Metalloproteinases are largely responsible for degradation of the basement membrane of microvessels, with the consequent hemorrhage (Escalante et al, 2011), activation of prothrombin and factor X (Kini, 2005;Tans & Rosing, 2001), thus generating the formation of microthrombi and fibrinogen depletion, i.e. defibrinogenation , and degradation of the extracellular matrix (Moura-da-Silva et al, 2009), among other effects.…”

Section: Snake Venom Biochemistry and Toxicologymentioning

confidence: 99%

Snakebite Envenoming: A Public Health Perspective

Gutiérrez¹

2012

Public Health - Methodology, Environmental and Systems Issues

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“…Por exemplo, SVMPs das classes P-II e P-III normalmente induzem hemorragia, e da classe P-I, não são hemorrágicas ou induzem muito pouca hemorragia; esta disparidade se deve basicamente à presença dos domínios externos dis-like e cys-rich (em P-III) e um tipo desintegrina (em P-II) na estrutura tridimensional destas metaloproteases, os quais facilmente se ligam aos componentes da membrana basal (BM) e a células endoteliais, danificando assim os vasos capilares, sem contar que estes domínios externos também se ligam a integrinas de plaquetas, ajudando desta forma para que não ocorra agregação plaquetária e, consequentemente contribuindo para o maior potencial hemorrágico (ESCALANTE et al, 2011;SAJEVIC et al, 2011). Enquanto isso, as metaloproteases P-I, apesar da pouca ou nenhuma atividade hemorrágica, têm revelado alta atividade proteolítica sobre os componentes da cascata de coagulação sanguínea, especialmente sobre o fibrinogênio (Fator I).…”

Section: Resultsunclassified

“…Nas reações inflamatórias, tem provável papel no reparo tecidual e na cicatrização, sem contar que a dosagem sanguínea do fibrinogênio permite aos profissionais da área de saúde identificar patologia relacionada à coagulação (FOX; SERRANO, 2010;SAJEVIC et al, 2011).…”

Section: Resultsunclassified

“…As peçonhas botrópicas são, portanto, capazes de ativar fatores da coagulação sanguínea, ocasionando o consumo de fibrinogênio, formação de fibrina e induzindo frequentemente à incoagulabilidade sanguínea. A maioria das serpentes do gênero Bothrops apresenta, isolada ou simultaneamente, componentes capazes de ativar o fibrinogênio, a protombina e outros fatores da cascata de coagulação, sobretudo FII, FV e FX (ESCALANTE et al, 2011;SAJEVIC et al, 2011;MATSUI et al, 2000).…”

Section: Introductionunclassified

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Purificação e avaliação da interferência de BthMP (uma metaloprotease da peçonha de Bothrops moojeni) na coagulação sanguínea

et al. 2016

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In the present was work reported the purification of the metalloprotease BthMP, from the venom of Bothrops moojeni. For purification of the protease was used ion exchange chromatography (DEAE-Sepharose) and molecular exclusion , the product of these processes was a protein band with high purity, visualized on SDS-PAGE 14%, referred the BthMP. This protease when analyzed on MALDI-TOF revealed the molecular weight of the native form of 23.050 Da and 23.872 Da in the reduced form, and from the peptide fragments obtained by Peptide Mass Fingerprinting (PMF) in MS (MALDI-TOF/TOF) was observed high similarity with BmooMPα-I metalloprotease. In enzymatic terms, BthMP showed proteolytic activity on azocasein using PMSF and benzamidine, while this activity was inhibited in the presence of EDTA; 1,10-Phenanthroline and β-mercaptoethanol, it is therefore a zinc-dependent metalloprotease of the class P-I. To still with this purpose, we contemplate its enzymatic specificity of the Aα and Bβ chains of fibrinogen and also the consumption of fibrinogen in vivo. It was also found its action on the components of the coagulation cascade, due to prolongation of prothrombin time and partial thromboplastin time. Thus, the sharp fibrinogenolytic activity and the high consumption of fibrinogen in vivo are results that indicate the anticoagulant action of BthMP; furthermore, their ability to interfere with the coagulation cascade suggested that this protease is promising for future studies that might indicate a new antithrombotic agent model. K E Y W O R D SCoagulation cascade, Metalloproteinase, Venom snake R E S U M ONeste trabalho relatamos a purificação da metaloprotease BthMP, proveniente da peçonha da serpente Bothrops moojeni. Para a purificação desta protease, utilizaram-se os passos cromatográficos de troca iônica (DEAE-Sepharose) e de exclusão molecular (Sephadex G-75), sendo o produto desses processos uma banda proteica com elevado grau de pureza, visualizada em SDS-PAGE a 14%, denominada BthMP. Esta, por sua vez, quando analisada em MALDI-TOF revelou a massa molecular nativa de 23.050 Da e 23.872 Da na forma reduzida, e a partir dos fragmentos peptídicos obtidos por Peptide Mass Fingerprinting (PMF) em MS (MALDI-TOF/TOF) indicou alta similaridade com a metaloprotease BmooMPα-I. Em termos enzimáticos, BthMP mostrou atividade proteolítica sobre azocaseína e frente ao PMSF e benzamidina, enquanto que esta atividade foi inibida na presença de EDTA, 1,10-fenantrolina e β-mercaptoetanol, sendo portanto uma metaloprotease zinco dependente da classe P-I. Ainda com este propósito, verificou-se sua especificidade enzimática sobre as cadeias Aα e Bβ do fibrinogênio, e também o consumo de fibrinogênio in vivo. Foi constatado ainda sua ação em componentes da cascata de coagulação, devido ao prolongamento do Tempo de Protrombina (TP) e do Tempo de Tromboplastina Parcial ativada (TTPa). Desta forma, a acentuada atividade fibrinogenolítica e o alto consumo de fibrinogênio in vivo são resultados que indicam a ação anticoagulante da BthMP; al...

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Key events in microvascular damage induced by snake venom hemorrhagic metalloproteinases

Cited by 208 publications

References 89 publications

Antibodies against Venom of the Snake Deinagkistrodon acutus

Antibodies against Venom of the Snake Deinagkistrodon acutus

Snakebite Envenoming: A Public Health Perspective

Purificação e avaliação da interferência de BthMP (uma metaloprotease da peçonha de Bothrops moojeni) na coagulação sanguínea

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