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Nadia C G de Morais,
Carla C Neves Mamede,
Kelly C Fonseca, Mayara R de Queiroz,
Saulo A Gomes-Filho,
Norival A Santos-Filho,
Karla de C F Bordon,
Marcelo E Beletti,
Suely V Sampaio,
Eliane C Arantes,
Fábio de Oliveira
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ABSTRACT: A fibrinogenolytic metalloproteinase from Bothrops moojeni venom, named moojenin, was purified by a combination of ion-exchange chromatography on DEAE-Sephacel and gel filtration on Sephacryl S-300. SDS-PAGE analysis indicated that moojenin consists of a single polypeptide chain and has a molecular mass about 45 kDa. Sequencing of moojenin by Edman degradation revealed the amino acid sequence LGPDIVSPPVCGNELLEVGEECDCGTPENCQNE, which showed strong identity with many other snake venom metalloproteinases (SVMPs). The enzyme cleaves the Aα-chain of fibrinogen first, followed by the Bβ-chain, and shows no effects on the γ-chain. Moojenin showed a coagulant activity on bovine plasma about 3.1 fold lower than crude venom. The fibrinogenolytic and coagulant activities of the moojenin were abolished by preincubation with EDTA, 1,10-phenanthroline and β-mercaptoethanol. Moojenin showed maximum activity at temperatures ranging from 30 to 40 °C and its optimal pH was 4.0. Its activity was completely lost at temperatures above 50 °C. Moojenin induced necrosis in liver and muscle, evidenced by morphological alterations, but did not cause histological alterations in mouse lungs, kidney or heart. Moojenin rendered the blood uncoagulatable when it was intraperitoneally administered into mice. This metalloproteinase may be of medical interest because of its anticoagulant activity.
Toxicon 09/2012; 60(7):1251-8. · 2.51 Impact Factor
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Mário Sérgio R Gomes, Mayara R de Queiroz,
Carla C N Mamede,
Mirian M Mendes,
Amélia Hamaguchi,
Maria I Homsi-Brandeburgo,
Marcelo V Sousa,
Elaine Nascimento Aquino,
Mariana S Castro,
Fábio de Oliveira,
Veridiana M Rodrigues
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ABSTRACT: A fibrino(geno)lytic nonhemorrhagic metalloproteinase (BleucMP) was purified from Bothrops leucurus snake venom by two chromatographic steps procedure on DEAE-Sephadex A-25 followed by CM-Sepharose Fast Flow column. BleucMP represented 1.75% (w/w) of the crude venom and was homogeneous on SDS-PAGE. BleucMP analyzed by MALDI TOF/TOF, showed a molecular mass of 23,057.54Da and when alkylated and reduced, the mass is 23,830.40Da. Their peptides analyzed in MS (MALDI TOF\TOF) showed significant score when compared with those of other proteins by NCBI-BLAST2 alignment display. As regards their proteolytic activities, BleucMP efficiently acted on fibrinogen, fibrin, and was inhibited by EDTA and 1.10-phenanthroline. This enzyme was also able to decrease significantly the plasma fibrinogen level provoking blood incoagulability, however was devoid of hemorrhagic activity when tested in the mice skin and did not induce relevant biochemical, hematological and histopathological alterations in mice. The aspects addressed in this paper provide data on the effect of BleucMP in envenomation from B. leucurus snakes in order to better understand the effects caused by snake venom metalloproteinase.
Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology 04/2011; 153(3):290-300. · 2.62 Impact Factor
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Mário Sérgio R. Gomes, Mayara R. de Queiroz,
Carla C.N. Mamede,
Mirian M. Mendes,
Amélia Hamaguchi,
Maria I. Homsi-Brandeburgo,
Marcelo V. Sousa,
Elaine Nascimento Aquino,
Mariana S. Castro,
Fábio de Oliveira,
Veridiana M. Rodrigues
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[hide abstract]
ABSTRACT: A fibrino(geno)lytic nonhemorrhagic metalloproteinase (BleucMP) was purified from Bothrops leucurus snake venom by two chromatographic steps procedure on DEAE-Sephadex A-25 followed by CM-Sepharose Fast Flow column. BleucMP represented 1.75% (w/w) of the crude venom and was homogeneous on SDS-PAGE. BleucMP analyzed by MALDI TOF/TOF, showed a molecular mass of 23,057.54 Da and when alkylated and reduced, the mass is 23,830.40 Da. Their peptides analyzed in MS (MALDI TOF\TOF) showed significant score when compared with those of other proteins by NCBI-BLAST2 alignment display. As regards their proteolytic activities, BleucMP efficiently acted on fibrinogen, fibrin, and was inhibited by EDTA and 1.10-phenanthroline. This enzyme was also able to decrease significantly the plasma fibrinogen level provoking blood incoagulability, however was devoid of hemorrhagic activity when tested in the mice skin and did not induce relevant biochemical, hematological and histopathological alterations in mice. The aspects addressed in this paper provide data on the effect of BleucMP in envenomation from B. leucurus snakes in order to better understand the effects caused by snake venom metalloproteinase.
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology. 153(3):290-300.
