Poster 110.  Congresso da Sociedade Brasileira de Toxinologia, 8., Symposium of the Pan American Section of the International Society on Toxinology, 8., 2004, Angra dos Reis, Brasil.  Abstracts...  J. Venom. Anim. Toxins incl.Trop. Dis., 2004, 10, 3, p.469.

 

 

Functional and Enzymatic Properties of BjussuMP-I, a New Snake Venom Metalloprotease from Bothrops jararacussu. 

 

1Mazzi, M. V.; 2Marcussi, S.; 2Carlos, G. B.; 3Stábeli, R. G.; 1Ticli, F. K.; 1Silva, J. O.; 1Sant´Ana, C. D.; 2França, S. C.; 2Soares, A. M.; 1Sampaio, S. V.

 

1 Depto. Análises Clínicas, Toxicológicas e Bromatológicas, FCFRP/USP; 2 Unidade de Biotecnologia, UNAERP, Ribeirão Preto-SP; 3 IPEPATRO, Porto Velho-RO and 4 Depto. Bioquímica e Imunologia, FMRP/USP, Ribeirão Preto-SP, Brazil.

 

Features of Bothrops genus envenomation are characterized by serious muscle damage, hemorrhage and blood coagulation disorders. This effect arises due to synergic action of proteolytic enzymes, such as different class of metalloproteases. From the venom of snake Bothrops jararacusssu a hemorrhagic metalloprotease, named BjussuMP-I, is a 60kDa protein with a pI~5.5, which induced hemorrhage in mice (MHD=4.0 µg). Hemorrhagic activity of BjussuMP-I was totally abolished after incubation with chelating agent (EDTA), corroborating the metal-dependency of this effect. Despite exhibiting hemorrhagic activity, myotoxic activity is null when compaired with a myotoxic PLA2 (BthTX-I). The enzyme is able to inhibit human platelet aggregation in a dose-dependent manner. BjussuMP-I shows proteolytic activity on casein, fibrinogen and fibrin, hydrolyzing A chain of fibrinogen more efficiently than B chain and is active in acid and basic conditions, however was inactivated at both 4 and 75°C. The A chain of fibrinogen was completely hydrolised in the presence of Ca++ and Mg++, although some ions (Mn++, Fe++, Zn++, Co++ and Ni++) shows a lower influence on fibrinogen degradation. This activity is completely inhibted by EDTA, EGTA and partially by PMSF. Fibrinolytic activity is observed with BjussuMP-I (0.2 mg/mL) at 37°C for 18-24 h. BjussuMP-I also shows bactericidal activity against Escherichia coli and Staphylococcus aureus. This first report on the isolation and characterization of a new high molecular weight hemorrhagic metalloprotease (BjussuMP-I) may play a relevant role in local and systemic bleeding characteristic of Bothrops envenomations, and prove to be invaluable pharmacological probe.

 

Support: FAPESP, CNPq and UNAERP.

 

Correspondence to: maumazzi@fcfrp.usp.br, andreimar@unaerp.br or suvilela@fcfrp.usp.br.