Isolation and structural characterization of a new fibrin(ogen)olytic metalloproteinase from Bothrops moojeni snake venom

被引：60

作者：

Bernardes, Carolina P. ^{[1
,2
]}

Santos-Filho, Norival A. ^{[1
,2
]}

Costa, Tassia R. ^{[1
]}

Gornes, Mario S. R. ^{[3
,4
]}

Torres, Fernanda S. ^{[2
]}

Costa, Junia ^{[2
]}

Borges, Marcia H. ^{[5
]}

Richardson, Michael ^{[5
]}

dos Santos, Daniel M. ^{[6
]}

de Castro Pimenta, Adriano M. ^{[6
]}

Homsi-Brandeburgo, Maria I. ^{[2
]}

Soares, Andreimar M. ^{[1
]}

de Oliveira, Fabio ^{[2
]}

机构：

[1] Univ Sao Paulo, FCFRP USP, Fac Ciencias Farmaceut, Dept Anal Clin Toxicol & Bromatol, BR-14049 Ribeirao Preto, Brazil

[2] Univ Fed Uberlandia, Inst Ciencias Biomed, BR-38400902 Uberlandia, MG, Brazil

[3] Univ Fed Uberlandia, Inst Genet & Bioquim, BR-38400902 Uberlandia, MG, Brazil

[4] Univ Estadual Sudoeste Bahia, Dept Quim & Exatas, BR-45506210 Jequie, BA, Brazil

[5] Fundacao Ezequiel Dias, Diretoria Pesquisa & Desenvolvimento, BR-30510010 Belo Horizonte, MG, Brazil

[6] Univ Fed Minas Gerais, Dept Bioquim & Imunol, BR-31270901 Belo Horizonte, MG, Brazil

来源：

TOXICON | 2008年 / 51卷 / 04期

基金：

巴西圣保罗研究基金会;

关键词：

Bothrops moojeni; snake venom; metalloproteinase; fibrinogenase; structural characterization;

D O I：

10.1016/j.toxicon.2007.11.017

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

A proteinase, named BmooMP alpha-I, from the venom of Bothrops moojeni, was purified by DEAE-Sephacel, Sephadex G-75 and heparin-agarose column chromatography. The enzyme was purified to homogeneity as judged by its migration profile in SDS-PAGE stained with coomassie blue, and showed a molecular mass of about 24.5 kDa. Its complete cDNA was obtained by RT-PCR and the 615 bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteinases showed a high structural similarly, mainly among class P-IB proteases. The enzyme cleaves the A alpha-chain of fibrinogen first, followed by the B beta-chain, and shows no effects on the gamma-chain. On fibrin, the enzyme hydrolyzed only the beta-chain, leaving the gamma-dimer apparently untouched. It was devoid of phospholipase A(2), hemorrhagic and thrombin-like activities. Like many venom enzymes, it is stable at pH values between 4 and 10 and stable at 70 degrees C for 15 min. The inhibitory effects of EDTA on the fibrinogenolytic activity suggest that BmooMP alpha-I is a metalloproteinase and inhibition by beta-mercaptoethanol revealed the important role of the disulfide bonds in the stabilization of the native structure. Aprotinin and benzamidine, specific serine proteinase inhibitors, had no effect on BmooMP alpha-I activity. Since the BmooMP alpha-I enzyme was found to cause defibrinogenation when administered i.p. on mice, it is expected that it may be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis. (C) 2007 Elsevier Ltd. All rights reserved.

引用

页码：574 / 584

页数：11

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[21] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].