Purification and biochemical and functional characterization of fibrinogen from plasma of Bothrops jararaca (Wied, 1824) (Ophidia: Viperidae, Crotalinae) / Purificação e caracterização bioquímica e funcional do fibrinogênio do plasma da serpente Bothrops jararaca (Wied, 1824) (Ophidia: Viperidae, Crotalinae)

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

Fibrinogen is a plasma glycoprotein that is composed of two sets of three nonidentical polypeptide chains (Aα Bβ and ϒ ) which are covalently linked by disulfide bonds. Thrombin cleaves fibrinogen to form fibrin and, consequently, the fibrin clot. This work describes the purification and characterization of fibrinogen from Bothrops jararaca (B. jararaca) snake plasma. Purified fibrinogen was obtained by barium chloride adsorption, ammonium sulfate precipitation and gel filtration chromatography. B. jararaca fibrinogen showed a molecular mass of 370 kDa in non-reducing conditions, similar to human and bovine fibrinogen with 340 kDa. Reduced fibrinogen showed three chains of 71, 60 and 55 kDa, which are similar to the molecular masses of human and bovine Aα Bβ and ϒ fibrinogen chains (64, 56 and 47 kDa, respectively). B. jararaca fibrinogen was clotted by bovine thrombin, however, B. jararaca, Crotalus durissus terrificus and Lachesis muta rhombeata venoms were not able to induce fibrin formation. The N-terminal sequence of B. jararaca fibrinogen chains from PVDF membranes showed only the first eight amino acids residues from each chain. The Nterminal sequence was Gly-Asp-Pro-Glu-Asp-Tyr-Leu-Gly for Aα chain, Gly-Ser-Asp- His-Asp-Asp-Glu for Bβ chain, and Glu-Ser-X-Leu-Asp-Glu-Glu-Gly for ϒ chain. The B. jararaca fibrinogen chains N-terminal sequences were compared to other animal Nterminal sequences previously described. However, due to low signal detection during Edman degradation, the sequence results were not sufficient to provide an accurate Blast search identity (NCBI-National Center for Biotechnology Information, www.ncbi.nih.gov). Mass spectrometry (MALDI-TOF) analysis provides some peptide sequences that did not present the complete sequences. Besides, anti-B. jararaca fibrinogen produced in rabbit did not recognize human fibrinogen while anti-human fibrinogen recognized B. jararaca fibrinogen. Thus, despite B. jararaca fibrinogen presents a molecular mass similar to human fibrinogen, the former shows distinctive features, which protect B. jararaca snakes from a fortuitous ingress of snake venom proteins into snake circulation, which could cause a self-envenomation

ASSUNTO(S)

plasma plasma coagulação sanguinea snake plasma plasma de serpente bothrops jararaca fibrinogen bothrops jararaca bood coagulation fibrinogênio

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