Purification and functional characterization of a new metalloproteinase (BleucMP) from Bothrops leucurus snake venom

Gomes, Marcos Antônio Silvestre; Queiroz, Mayara Ribeiro de; Mamede, Carla Cristine Neves; Mendes, Mirian Machado; Hamaguchi, Amélia; Homsi-Brandeburgo, Maria Inês; Sousa, Marcelo Valle de; Aquino, Elaine Nascimento; Castro, Mariana S.; Oliveira, Fábio de; Rodrigues, Veridiana M.

doi:10.1016/j.cbpc.2010.11.008

Cited by 30 publications

(17 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Electrophoretic analysis (SDS-PAGE) under reducing and nonreducing conditions indicated that the BmooMP α -II enzyme had a molecular mass about 25.5 and 22.5 kDa, respectively (Figure 1(c)). This molecular mass is similar to other bothropic SVMPs such as BjussuMP-II (24 kDa) from B. jararacussu [26], BmooMP α -I (24.5 kDa) from B. moojeni [27], Atroxlysin-I (23 kDa) from B. atrox [28], and BleucMP (23.5 kDa) from B. leucurus [10]. …”

Section: Resultssupporting

confidence: 74%

See 1 more Smart Citation

Rapid Purification of a New P-I Class Metalloproteinase fromBothrops moojeniVenom with Antiplatelet Activity

Mamede

Fonseca

et al. 2014

BioMed Research International

Self Cite

View full text Add to dashboard Cite

The present study aimed to evaluate the proteolytic and biological activities of a new metalloproteinase from B. moojeni venom. The purification of BmooMPα-II was carried out through two chromatographic steps (ion-exchange and affinity). BmooMPα-II is a monomeric protein with an apparent molecular mass of 22.5 kDa on SDS-PAGE 14% under nonreducing conditions. The N-terminal sequence (FSPRYIELVVVADHGMFTKYKSNLN) revealed homology with other snake venom metalloproteinases, mainly among P-I class. BmooMPα-II cleaves Aα-chain of fibrinogen followed by Bβ-chain, and does not show any effect on the γ-chain. Its optimum temperature and pH for the fibrinogenolytic activity were 30–50°C and pH 8, respectively. The inhibitory effects of EDTA and 1,10-phenantroline on the fibrinogenolytic activity suggest that BmooMPα-II is a metalloproteinase. This proteinase was devoid of haemorrhagic, coagulant, or anticoagulant activities. BmooMPα-II caused morphological alterations in liver, lung, kidney, and muscle of Swiss mice. The enzymatically active protein yet inhibited collagen, ADP, and ristocetin-induced platelet aggregation in a concentration-dependent manner. Our results suggest that BmooMPα-II contributes to the toxic effect of the envenomation and that more investigations to elucidate the mechanisms of inhibition of platelet aggregation may contribute to the studies of snake venom on thrombotic disorders.

show abstract

Section: Resultssupporting

confidence: 74%

“…A α - and B β -chain degradation was completed after 60 min incubation (Figure 3(a)). Therefore, BmooMP α -II is classified as α -fibrinogenase, similar to neuwiedase [23], BlaH1 [29], BjussuMP-II [26], BmooMP α -I [27], BthMP [9], and BleucMP [10]. …”

Section: Resultsmentioning

confidence: 99%

Rapid Purification of a New P-I Class Metalloproteinase fromBothrops moojeniVenom with Antiplatelet Activity

Mamede

Fonseca

et al. 2014

BioMed Research International

Self Cite

View full text Add to dashboard Cite

show abstract

“…Esta massa molecular condiz com o já estimado anteriormente em SDS-PAGE 14%, que foi em torno de 23 kDa (GOMES et al, 2011), porém em um nível maior de precisão devido a sensibilidade da técnica utilizada. A forma reduzida desta proteína foi obtida através do uso de agente redutor, o qual tem a característica de romper ligações dissulfeto da estrutura terciária da proteína.…”

Section: Resultsunclassified

“…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). Muitas das SVMPs da classe P-I apresentam este caráter, das quais temos BJ-P12 de B. jararaca (SILVA et al, 2012); Batroxase de B. atrox (CINTRA et al, 2012); BleucMP de B. leucurus (GOMES et al, 2011); dentre outras. Estas metaloproteases são chamadas de enzimas fibrinogenolíticas, porque provocam o consumo do fibrinogênio, causando assim uma desfibrin (ogen)ação e, por consequência, tornando o sangue mais fluido e aumentando o tempo de coagulação.…”

Section: Resultsunclassified

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

et al. 2016

View full text Add to dashboard Cite

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...

show abstract

“…jararacussu (Marcussi et al, 2007), BmooMPα-I from B. moojeni (Bernardes et al, 2008), and BleucMP from B. leucurus (Gomes et al, 2011) are some examples of α-fibrinogenases described in the literature. The inhibition of platelet aggregation caused by SVMPs could be attributed to degradation of fibrinogen, the main ligand of the α IIb β 3 integrin.…”

Section: A C C E P T E D Accepted Manuscriptmentioning

confidence: 97%

The role of platelets in hemostasis and the effects of snake venom toxins on platelet function

Queiroz

Sousa

Pereira

et al. 2017

Toxicon

Self Cite

View full text Add to dashboard Cite

Purification and functional characterization of a new metalloproteinase (BleucMP) from Bothrops leucurus snake venom

Cited by 30 publications

References 43 publications

Rapid Purification of a New P-I Class Metalloproteinase fromBothrops moojeniVenom with Antiplatelet Activity

Rapid Purification of a New P-I Class Metalloproteinase fromBothrops moojeniVenom with Antiplatelet Activity

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

The role of platelets in hemostasis and the effects of snake venom toxins on platelet function

Contact Info

Product

Resources

About